mcherry plasmid  (New England Biolabs)


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    Structured Review

    New England Biolabs mcherry plasmid
    NvLsd1 is required cell autonomously for cnidocyte differentiation ( A-C ) Immunostaining on NvLsd1 −/− animals injected with NvPOU4 <t>:NvLsd1-mCherry</t> ( A ), NvPOU4 :NvH2B-mCherry ( B ) or NvPOU4 :NvLsd1 K644A/A520E -mCherry ( C ). DNA is shown in blue, DAPI + cnidocysts in green and mCherry in magenta. Scale bars: 10 μm.
    Mcherry Plasmid, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mcherry plasmid/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mcherry plasmid - by Bioz Stars, 2022-07
    86/100 stars

    Images

    1) Product Images from "Histone demethylase Lsd1 is required for the differentiation of neural cells in the cnidarian Nematostella vectensis"

    Article Title: Histone demethylase Lsd1 is required for the differentiation of neural cells in the cnidarian Nematostella vectensis

    Journal: bioRxiv

    doi: 10.1101/2020.09.07.285577

    NvLsd1 is required cell autonomously for cnidocyte differentiation ( A-C ) Immunostaining on NvLsd1 −/− animals injected with NvPOU4 :NvLsd1-mCherry ( A ), NvPOU4 :NvH2B-mCherry ( B ) or NvPOU4 :NvLsd1 K644A/A520E -mCherry ( C ). DNA is shown in blue, DAPI + cnidocysts in green and mCherry in magenta. Scale bars: 10 μm.
    Figure Legend Snippet: NvLsd1 is required cell autonomously for cnidocyte differentiation ( A-C ) Immunostaining on NvLsd1 −/− animals injected with NvPOU4 :NvLsd1-mCherry ( A ), NvPOU4 :NvH2B-mCherry ( B ) or NvPOU4 :NvLsd1 K644A/A520E -mCherry ( C ). DNA is shown in blue, DAPI + cnidocysts in green and mCherry in magenta. Scale bars: 10 μm.

    Techniques Used: Immunostaining, Injection

    2) Product Images from "EFF-1 promotes muscle fusion, paralysis and retargets infection by AFF-1-coated viruses in C. elegans"

    Article Title: EFF-1 promotes muscle fusion, paralysis and retargets infection by AFF-1-coated viruses in C. elegans

    Journal: bioRxiv

    doi: 10.1101/2020.05.17.099622

    EFF-1 expression in BWM enables VSVΔG-AFF-1 and VSVΔG-G spreading along fused muscles (A-L) Z-stack projections of wt-like (A-F) and Unc+Dpy animals (G-L) expressing myo-3p:: EFF-1 and myo-3p:: mCherry infected with VSVΔG-AFF-1 (35-63 IU red pins) or VSVΔG-G (3*10 5 IU; white pins). Insets and their corresponding images (yellow frames). Arrows, individual infected (cyan) BWMs. Dashed lines outline grouped BWMs that express myo-3p ::mCherry and myo-3p:: EFF-1 (magenta) and infected with virus (cyan) showing spreading of GFP. Scale bars, 100 µm. (M-N) Number of BWM cells/worm expressing EFF-1 (magenta cell), infected (cyan cell) or expressing EFF-1 and infected (magenta and cyan). wt-like (circles) and Unc+Dpy (triangles). Each point represents a single worm. (O-P) Quantitation of multinucleation of infected BWMs. Each dot represents an average number of nuclei/ GFP(+) BWM, calculated from 1-6 multinucleated BWMs of a single worm. (M and O) wt-like n=6 and Unc+Dpy n=10 animals. (N and P) wt-like n=9 and Unc+Dpy n=7 animals. Black horizontal lines, average ± SEM. Student’s t-test, *** p
    Figure Legend Snippet: EFF-1 expression in BWM enables VSVΔG-AFF-1 and VSVΔG-G spreading along fused muscles (A-L) Z-stack projections of wt-like (A-F) and Unc+Dpy animals (G-L) expressing myo-3p:: EFF-1 and myo-3p:: mCherry infected with VSVΔG-AFF-1 (35-63 IU red pins) or VSVΔG-G (3*10 5 IU; white pins). Insets and their corresponding images (yellow frames). Arrows, individual infected (cyan) BWMs. Dashed lines outline grouped BWMs that express myo-3p ::mCherry and myo-3p:: EFF-1 (magenta) and infected with virus (cyan) showing spreading of GFP. Scale bars, 100 µm. (M-N) Number of BWM cells/worm expressing EFF-1 (magenta cell), infected (cyan cell) or expressing EFF-1 and infected (magenta and cyan). wt-like (circles) and Unc+Dpy (triangles). Each point represents a single worm. (O-P) Quantitation of multinucleation of infected BWMs. Each dot represents an average number of nuclei/ GFP(+) BWM, calculated from 1-6 multinucleated BWMs of a single worm. (M and O) wt-like n=6 and Unc+Dpy n=10 animals. (N and P) wt-like n=9 and Unc+Dpy n=7 animals. Black horizontal lines, average ± SEM. Student’s t-test, *** p

    Techniques Used: Expressing, Infection, Quantitation Assay

    EFF-1 expression in BWMs produces Unc+Dpy worms with multinucleated cells (A-H) Images of fluorescent Z-stack projections and respective DIC of animals with extrachromosomal myo-3p ::EFF-1, myo-3p ::mCherry. (G) White arrowhead, bridge formed between 2 BWMs from opposing quadrants. Yellow arrows, myo-3p ::mCherry accumulations also in DIC (H). Asterisks, clustered nuclei within one BWM. Scale bars, 100 µm. (I) Number of nuclei per myo-3p ::mCherry (+) BWM cell in L2s. wt-like (n=12); Unc+Dpy (n=15). Each dot represents the average number of nuclei/BWM cell/worm. Total average ± SEM for each phenotype. Two tailed Student’s t-test p
    Figure Legend Snippet: EFF-1 expression in BWMs produces Unc+Dpy worms with multinucleated cells (A-H) Images of fluorescent Z-stack projections and respective DIC of animals with extrachromosomal myo-3p ::EFF-1, myo-3p ::mCherry. (G) White arrowhead, bridge formed between 2 BWMs from opposing quadrants. Yellow arrows, myo-3p ::mCherry accumulations also in DIC (H). Asterisks, clustered nuclei within one BWM. Scale bars, 100 µm. (I) Number of nuclei per myo-3p ::mCherry (+) BWM cell in L2s. wt-like (n=12); Unc+Dpy (n=15). Each dot represents the average number of nuclei/BWM cell/worm. Total average ± SEM for each phenotype. Two tailed Student’s t-test p

    Techniques Used: Expressing, Two Tailed Test

    VSVΔG-AFF-1 does not infect PVD and other sensory neurons ectopically expressing AFF-1/EFF-1 (A-C) SDC microscope Z-stack projections of animals infected with 82-103 IU VSVΔG-AFF-1 (red pins). (For genotypes and quantitation see Table S2). Scale bar, 100 µm. (A) Young adult expressing mCherry in PVD. (B) eff-1(ts) adult expressing AFF-1 in PVD. (C) eff-1(ts) adult expressing EFF-1 and dsRed in 12 sensory neurons. See also Table S2 and Movie S2.
    Figure Legend Snippet: VSVΔG-AFF-1 does not infect PVD and other sensory neurons ectopically expressing AFF-1/EFF-1 (A-C) SDC microscope Z-stack projections of animals infected with 82-103 IU VSVΔG-AFF-1 (red pins). (For genotypes and quantitation see Table S2). Scale bar, 100 µm. (A) Young adult expressing mCherry in PVD. (B) eff-1(ts) adult expressing AFF-1 in PVD. (C) eff-1(ts) adult expressing EFF-1 and dsRed in 12 sensory neurons. See also Table S2 and Movie S2.

    Techniques Used: Expressing, Microscopy, Infection, Quantitation Assay

    EFF-1 expression in BWMs induces their fusion ( A-C ) Confocal images of wt-like adult worms with membrane bound ( MB ) myo-3p::MB::YFP (cyan) and extrachromosomal array containing myo-3p:: EFF-1, myo-3p:: mCherry (magenta). ( D-F ) Confocal images of Unc+Dpy [ myo-3p::MB::YFP (cyan); myo-3p::EFF-1, myo-3p::mCherry] . Arrows, unfused BWMs with MB (cyan). Note only two unfused BWMs, all the others appear fused with no MB separating them. Insets correspond to white-dotted area. Scale bars 100 µm. See also Movie S1.
    Figure Legend Snippet: EFF-1 expression in BWMs induces their fusion ( A-C ) Confocal images of wt-like adult worms with membrane bound ( MB ) myo-3p::MB::YFP (cyan) and extrachromosomal array containing myo-3p:: EFF-1, myo-3p:: mCherry (magenta). ( D-F ) Confocal images of Unc+Dpy [ myo-3p::MB::YFP (cyan); myo-3p::EFF-1, myo-3p::mCherry] . Arrows, unfused BWMs with MB (cyan). Note only two unfused BWMs, all the others appear fused with no MB separating them. Insets correspond to white-dotted area. Scale bars 100 µm. See also Movie S1.

    Techniques Used: Expressing

    Retargeting of VSVΔG-AFF-1 to body wall muscle cells Wild-type worms and animals with extrachromosomal array containing myo-3p:: EFF-1 and myo-3p:: mCherry were injected with VSVΔG-AFF-1 (35-63 IU, red pins; n=39 wt, n=50 wt-like and n=27 Unc+Dpy worms) or VSVΔG-G (3*10 5 IU, white pins; n=30 wt-like and 14 Unc+Dpy) respectively. Wt worms injected with VSVΔG-G (2300-4700 IU, n=56) were taken from figure 2I. Animals were analysed by SDC microscopy. Data represents average percentage of worms with GFP(+) BWMs ± SEM. Student’s t-test: *p
    Figure Legend Snippet: Retargeting of VSVΔG-AFF-1 to body wall muscle cells Wild-type worms and animals with extrachromosomal array containing myo-3p:: EFF-1 and myo-3p:: mCherry were injected with VSVΔG-AFF-1 (35-63 IU, red pins; n=39 wt, n=50 wt-like and n=27 Unc+Dpy worms) or VSVΔG-G (3*10 5 IU, white pins; n=30 wt-like and 14 Unc+Dpy) respectively. Wt worms injected with VSVΔG-G (2300-4700 IU, n=56) were taken from figure 2I. Animals were analysed by SDC microscopy. Data represents average percentage of worms with GFP(+) BWMs ± SEM. Student’s t-test: *p

    Techniques Used: Injection, Microscopy

    3) Product Images from "Histone demethylase Lsd1 is required for the differentiation of neural cells in Nematostella vectensis"

    Article Title: Histone demethylase Lsd1 is required for the differentiation of neural cells in Nematostella vectensis

    Journal: Nature Communications

    doi: 10.1038/s41467-022-28107-z

    NvLsd1 is required cell autonomously for cnidocyte differentiation. a – c Immunostaining on NvLsd1 −/− animals injected with NvPOU4 : NvLsd1-mCherry (n = 9) ( a ), NvPOU4 : NvH2B-mCherry (n = 16) ( b ) or NvPOU4 : NvLsd1 K644A/A520E -mCherry (n = 6) ( c ). DNA is shown in blue, DAPI + cnidocysts in green and mCherry in magenta. Experiments were performed three times independently and the total number of animals analysed for each condition are indicated (n). Scale bars: 10 μm.
    Figure Legend Snippet: NvLsd1 is required cell autonomously for cnidocyte differentiation. a – c Immunostaining on NvLsd1 −/− animals injected with NvPOU4 : NvLsd1-mCherry (n = 9) ( a ), NvPOU4 : NvH2B-mCherry (n = 16) ( b ) or NvPOU4 : NvLsd1 K644A/A520E -mCherry (n = 6) ( c ). DNA is shown in blue, DAPI + cnidocysts in green and mCherry in magenta. Experiments were performed three times independently and the total number of animals analysed for each condition are indicated (n). Scale bars: 10 μm.

    Techniques Used: Immunostaining, Injection

    4) Product Images from "Tissue-specific delivery system via AFF-1-coated pseudotyped Vesicular Stomatitis Virus in C. elegans"

    Article Title: Tissue-specific delivery system via AFF-1-coated pseudotyped Vesicular Stomatitis Virus in C. elegans

    Journal: bioRxiv

    doi: 10.1101/2020.05.17.099622

    EFF-1 in BWM enables VSVΔG-AFF-1 and VSVΔG-G spreading along fused muscles (A-L) Z-stack projections of wt-like (A-F) and Unc+Dpy animals (G-L) expressing myo-3p:: EFF-1 and myo-3p:: mCherry infected with VSVΔG-AFF-1 (35-63 IU red pins) or VSVΔG-G (3*10 5 IU; white pins). Insets and their corresponding images (yellow frames). Arrows, individual infected (cyan) BWMs. Dashed lines outline grouped BWMs that express myo-3p ::mCherry and myo-3p:: EFF-1 (magenta) and infected with virus (cyan) showing spreading of GFP. Scale bars, 100 µm. (M-N) Number of BWM cells/worm expressing EFF-1 (magenta cell), infected (cyan cell) or expressing EFF-1 and infected (magenta and cyan). wt-like (circles) and Unc+Dpy (triangles). Each point represents a single worm. (O-P) Quantitation of multinucleation of infected BWMs. Each dot represents an average number of nuclei/ GFP(+) BWM, calculated from 1-6 multinucleated BWMs of a single worm. (M and O) wt-like n=6 and Unc+Dpy n=10 animals. (N and P) wt-like n=9 and Unc+Dpy n=7 animals. Black horizontal lines, average ± SEM. Student’s t-test, *** p
    Figure Legend Snippet: EFF-1 in BWM enables VSVΔG-AFF-1 and VSVΔG-G spreading along fused muscles (A-L) Z-stack projections of wt-like (A-F) and Unc+Dpy animals (G-L) expressing myo-3p:: EFF-1 and myo-3p:: mCherry infected with VSVΔG-AFF-1 (35-63 IU red pins) or VSVΔG-G (3*10 5 IU; white pins). Insets and their corresponding images (yellow frames). Arrows, individual infected (cyan) BWMs. Dashed lines outline grouped BWMs that express myo-3p ::mCherry and myo-3p:: EFF-1 (magenta) and infected with virus (cyan) showing spreading of GFP. Scale bars, 100 µm. (M-N) Number of BWM cells/worm expressing EFF-1 (magenta cell), infected (cyan cell) or expressing EFF-1 and infected (magenta and cyan). wt-like (circles) and Unc+Dpy (triangles). Each point represents a single worm. (O-P) Quantitation of multinucleation of infected BWMs. Each dot represents an average number of nuclei/ GFP(+) BWM, calculated from 1-6 multinucleated BWMs of a single worm. (M and O) wt-like n=6 and Unc+Dpy n=10 animals. (N and P) wt-like n=9 and Unc+Dpy n=7 animals. Black horizontal lines, average ± SEM. Student’s t-test, *** p

    Techniques Used: Expressing, Infection, Quantitation Assay

    EFF-1 ectopic expression in BWMs results in Uncoordinated and Dumpy (Unc+Dpy) phenotypes Mixed population of worms with extrachromosomal pmyo-3::mCherry and pmyo-3::EFF-1 . White arrowhead point to mCherry(+) worm that is Unc+Dpy. White arrow point to mCherry(+) worm that is wt-like. Yellow arrowhead points to a wt-like mCherry(-) worm, which left the frame within seconds. Elapsed time (seconds) indicated in top left corner.
    Figure Legend Snippet: EFF-1 ectopic expression in BWMs results in Uncoordinated and Dumpy (Unc+Dpy) phenotypes Mixed population of worms with extrachromosomal pmyo-3::mCherry and pmyo-3::EFF-1 . White arrowhead point to mCherry(+) worm that is Unc+Dpy. White arrow point to mCherry(+) worm that is wt-like. Yellow arrowhead points to a wt-like mCherry(-) worm, which left the frame within seconds. Elapsed time (seconds) indicated in top left corner.

    Techniques Used: Expressing

    EFF-1 expression in BWMs produces Unc+Dpy worms with multinucleated cells (A-H) Images of fluorescent Z-stack projections and respective DIC of animals with extrachromosomal myo-3p ::EFF-1, myo-3p ::mCherry. (G) White arrowhead, bridge formed between 2 BWMs from opposing quadrants. Yellow arrows, myo-3p ::mCherry accumulations also in DIC (H). Asterisks, clustered nuclei within one BWM. Scale bars, 100 µm. (I) Number of nuclei per myo-3p ::mCherry (+) BWM cell in L2s. wt-like (n=12); Unc+Dpy (n=15). Each dot represents the average number of nuclei/BWM cell/worm. Total average ± SEM for each phenotype. Two tailed Student’s t-test p
    Figure Legend Snippet: EFF-1 expression in BWMs produces Unc+Dpy worms with multinucleated cells (A-H) Images of fluorescent Z-stack projections and respective DIC of animals with extrachromosomal myo-3p ::EFF-1, myo-3p ::mCherry. (G) White arrowhead, bridge formed between 2 BWMs from opposing quadrants. Yellow arrows, myo-3p ::mCherry accumulations also in DIC (H). Asterisks, clustered nuclei within one BWM. Scale bars, 100 µm. (I) Number of nuclei per myo-3p ::mCherry (+) BWM cell in L2s. wt-like (n=12); Unc+Dpy (n=15). Each dot represents the average number of nuclei/BWM cell/worm. Total average ± SEM for each phenotype. Two tailed Student’s t-test p

    Techniques Used: Expressing, Two Tailed Test

    VSVΔG-AFF-1 does not infect PVD and other sensory neurons ectopically expressing AFF-1/EFF-1 (A-C) SDC microscope Z-stack projections of animals infected with 82-103 IU VSVΔG-AFF-1 (red pins). (For genotypes and quantitation see Table S2 ). Scale bar, 100 µm. (A) Young adult expressing mCherry in PVD. (B) eff-1(ts) adult expressing AFF-1 in PVD. (C) eff-1(ts) adult expressing EFF-1 and dsRed in 12 sensory neurons. See also Table S2 and Movie S2 .
    Figure Legend Snippet: VSVΔG-AFF-1 does not infect PVD and other sensory neurons ectopically expressing AFF-1/EFF-1 (A-C) SDC microscope Z-stack projections of animals infected with 82-103 IU VSVΔG-AFF-1 (red pins). (For genotypes and quantitation see Table S2 ). Scale bar, 100 µm. (A) Young adult expressing mCherry in PVD. (B) eff-1(ts) adult expressing AFF-1 in PVD. (C) eff-1(ts) adult expressing EFF-1 and dsRed in 12 sensory neurons. See also Table S2 and Movie S2 .

    Techniques Used: Expressing, Microscopy, Infection, Quantitation Assay

    EFF-1 expression in BWMs induces their fusion ( A-C ) Confocal images of wt-like adult worms with membrane bound ( MB ) myo-3p::MB::YFP (cyan) and extrachromosomal array containing myo-3p:: EFF-1, myo-3p:: mCherry (magenta). ( D-F ) Confocal images of Unc+Dpy [ myo-3p::MB::YFP (cyan); myo-3p::EFF-1, myo-3p::mCherry] . Arrows, unfused BWMs with MB (cyan). Note only two unfused BWMs, all the others appear fused with no MB separating them. Insets correspond to white-dotted area. Scale bars 100 µm. See also Movie S1.
    Figure Legend Snippet: EFF-1 expression in BWMs induces their fusion ( A-C ) Confocal images of wt-like adult worms with membrane bound ( MB ) myo-3p::MB::YFP (cyan) and extrachromosomal array containing myo-3p:: EFF-1, myo-3p:: mCherry (magenta). ( D-F ) Confocal images of Unc+Dpy [ myo-3p::MB::YFP (cyan); myo-3p::EFF-1, myo-3p::mCherry] . Arrows, unfused BWMs with MB (cyan). Note only two unfused BWMs, all the others appear fused with no MB separating them. Insets correspond to white-dotted area. Scale bars 100 µm. See also Movie S1.

    Techniques Used: Expressing

    Retargeting of VSVΔG-AFF-1 to body wall muscle cells Wild-type worms and animals with extrachromosomal array containing myo-3p:: EFF-1 and myo-3p:: mCherry were injected with VSVΔG-AFF-1 (35-63 IU, red pins; n=39 wt, n=50 wt-like and n=27 Unc+Dpy worms) or VSVΔG-G (3*10 5 IU, white pins; n=30 wt-like and 14 Unc+Dpy) respectively. Wt worms injected with VSVΔG-G (2300-4700 IU, n=56) were taken from figure 2I . Animals were analysed by SDC microscopy. Data represents average percentage of worms with GFP(+) BWMs ± SEM. Student’s t-test: *p
    Figure Legend Snippet: Retargeting of VSVΔG-AFF-1 to body wall muscle cells Wild-type worms and animals with extrachromosomal array containing myo-3p:: EFF-1 and myo-3p:: mCherry were injected with VSVΔG-AFF-1 (35-63 IU, red pins; n=39 wt, n=50 wt-like and n=27 Unc+Dpy worms) or VSVΔG-G (3*10 5 IU, white pins; n=30 wt-like and 14 Unc+Dpy) respectively. Wt worms injected with VSVΔG-G (2300-4700 IU, n=56) were taken from figure 2I . Animals were analysed by SDC microscopy. Data represents average percentage of worms with GFP(+) BWMs ± SEM. Student’s t-test: *p

    Techniques Used: Injection, Microscopy

    5) Product Images from "Genomic integration and ligand-dependent activation of the human estrogen receptor α in the crustacean Daphnia magna"

    Article Title: Genomic integration and ligand-dependent activation of the human estrogen receptor α in the crustacean Daphnia magna

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0198023

    Structure of injected plasmids (A-C) and wild type Daphnia embryos injected with the plasmids at 50 ng/μL each and 26.8 ng/μL (100 μM) DES, at 16 hpi (D-I). (A) pRC21-hERa. Human estrogen receptor α ( esr1 ) with EF1α1 promoter and full length EF1α1 3’UTR. (B) pRC21-ERE:mCherry. Mcherry with 4 x ERE repeats and a truncated version of the EF1α1 3’UTR. (C) pRC21-EcRE:mCherry. Mcherry with 4 x EcRE repeats and a truncated version of the EF1α1 3’UTR. Lengths of the elements (not to scale) in bp are indicated underneath. (D) negative control (uninjected), (E) injection of pRC21-hERa and pRC21-ERE:mCherry, (F) injection of pRC21-ERE:mCherry and DES, (G) injection of pRC21-hERa and DES, (H) injection of pRC21-hERa, pRC21-ERE:mCherry and DES, (I) injection of control plasmid pRC21-EcRE:mCherry. Successful activation of the reporter is marked with white arrowheads. Bar = 100 μm.
    Figure Legend Snippet: Structure of injected plasmids (A-C) and wild type Daphnia embryos injected with the plasmids at 50 ng/μL each and 26.8 ng/μL (100 μM) DES, at 16 hpi (D-I). (A) pRC21-hERa. Human estrogen receptor α ( esr1 ) with EF1α1 promoter and full length EF1α1 3’UTR. (B) pRC21-ERE:mCherry. Mcherry with 4 x ERE repeats and a truncated version of the EF1α1 3’UTR. (C) pRC21-EcRE:mCherry. Mcherry with 4 x EcRE repeats and a truncated version of the EF1α1 3’UTR. Lengths of the elements (not to scale) in bp are indicated underneath. (D) negative control (uninjected), (E) injection of pRC21-hERa and pRC21-ERE:mCherry, (F) injection of pRC21-ERE:mCherry and DES, (G) injection of pRC21-hERa and DES, (H) injection of pRC21-hERa, pRC21-ERE:mCherry and DES, (I) injection of control plasmid pRC21-EcRE:mCherry. Successful activation of the reporter is marked with white arrowheads. Bar = 100 μm.

    Techniques Used: Injection, Negative Control, Plasmid Preparation, Activation Assay

    6) Product Images from "Genomic integration and ligand-dependent activation of the human estrogen receptor α in the crustacean Daphnia magna"

    Article Title: Genomic integration and ligand-dependent activation of the human estrogen receptor α in the crustacean Daphnia magna

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0198023

    Structure of injected plasmids (A-C) and wild type Daphnia embryos injected with the plasmids at 50 ng/μL each and 26.8 ng/μL (100 μM) DES, at 16 hpi (D-I). (A) pRC21-hERa. Human estrogen receptor α ( esr1 ) with EF1α1 promoter and full length EF1α1 3’UTR. (B) pRC21-ERE:mCherry. Mcherry with 4 x ERE repeats and a truncated version of the EF1α1 3’UTR. (C) pRC21-EcRE:mCherry. Mcherry with 4 x EcRE repeats and a truncated version of the EF1α1 3’UTR. Lengths of the elements (not to scale) in bp are indicated underneath. (D) negative control (uninjected), (E) injection of pRC21-hERa and pRC21-ERE:mCherry, (F) injection of pRC21-ERE:mCherry and DES, (G) injection of pRC21-hERa and DES, (H) injection of pRC21-hERa, pRC21-ERE:mCherry and DES, (I) injection of control plasmid pRC21-EcRE:mCherry. Successful activation of the reporter is marked with white arrowheads. Bar = 100 μm.
    Figure Legend Snippet: Structure of injected plasmids (A-C) and wild type Daphnia embryos injected with the plasmids at 50 ng/μL each and 26.8 ng/μL (100 μM) DES, at 16 hpi (D-I). (A) pRC21-hERa. Human estrogen receptor α ( esr1 ) with EF1α1 promoter and full length EF1α1 3’UTR. (B) pRC21-ERE:mCherry. Mcherry with 4 x ERE repeats and a truncated version of the EF1α1 3’UTR. (C) pRC21-EcRE:mCherry. Mcherry with 4 x EcRE repeats and a truncated version of the EF1α1 3’UTR. Lengths of the elements (not to scale) in bp are indicated underneath. (D) negative control (uninjected), (E) injection of pRC21-hERa and pRC21-ERE:mCherry, (F) injection of pRC21-ERE:mCherry and DES, (G) injection of pRC21-hERa and DES, (H) injection of pRC21-hERa, pRC21-ERE:mCherry and DES, (I) injection of control plasmid pRC21-EcRE:mCherry. Successful activation of the reporter is marked with white arrowheads. Bar = 100 μm.

    Techniques Used: Injection, Negative Control, Plasmid Preparation, Activation Assay

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    New England Biolabs mcherry ptp1b d181a
    Identification of <t>PTP1B</t> (also called PTPN1) in the BioID and as interactor of RHBDL4. A , peptide overage of PTP1B in the BioID of RHBDL4. Peptides identified by MS are shown in blue. B , the substrate-trapping mutant <t>D181A</t> and the super-trapping mutant D181A/Y46F of mCherry-PTP1B were transiently expressed with or without RHBDL4mycFLAG. The cells were lysed in immunoprecipitation ( IP )–compatible conditions and captured with anti-FLAG immunomagnetic beads. Co-immunoprecipitates and whole cell lysates ( Input ) were probed with anti-mCherry, or RHBDL4 antibody. C , HEK293 cells expressing TMEM115-mycFLAG or RHBDL4mycFLAG were lysed and bound with anti-FLAG beads ± 4 m m Na 3 VO 4 . The resulting co-immunoprecipitates were probed with anti-FLAG-HRP or anti-mCherry antibody.
    Mcherry Ptp1b D181a, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mcherry ptp1b d181a/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mcherry ptp1b d181a - by Bioz Stars, 2022-07
    86/100 stars
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    98
    New England Biolabs mcherry gene
    Phenotypic and genetic analysis of G 1 offspring. ( a ) Freshly hatched G 1 offspring, one <t>mCherry-negative</t> chicken in the middle. ( b ) Distribution of <t>mCherry-positive</t> cells in the peripheral blood. FACS histogram of mCherry signal in red blood cells (blue) and white blood cells (red). ( c ) mCherry positivity visible in the beak and featherless skin of the mCherry-positive rooster (right) in the daylight. The mCherry-negative wt rooster shown as a control (left). ( d ) Photostability of mCherry shown as mCherry signal in adult feather. ( e ) The 195 bp PCR product of the CB-specific tpn allele amplified in DNA of G 1 offspring. The results of animals Nos 8 to 14 (from left to right) are shown, the non-CB chicken No. 11 is in the middle. Descendants of G 1 roosters Nos 769 and 795 are indicated. ( f ) Genomic relationship of inbred individual CB145 with itself, inbred individual CB151, the primordial germ cell line, the mCherry+ chicken Robin and a mCherry- chicken. The genomic relationship was calculated based on about 1.6 million maximum quality SNP detected in 78 chickens.
    Mcherry Gene, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    New England Biolabs mcherry cenexin s796a plasmids
    Cenexin phosphorylation at its conserved C-terminal PLK1 binding site is required for maintenance of PCM in vivo . (A) Schematic representation of human hODF2 and cenexin (Cnxn, ODF2 isoform 9). The blue and magenta boxes highlight the N- and C-terminal extensions unique to cenexin. (B) Phylogenetic tree of the evolution of cenexin and its N-terminus and C-terminus in relation to Cep192 and centrin across different animal phyla. Cyan and orange boxes indicate whether cenexin and centrosome components (Cep192 and centrin) are detected or not detected within representative species of each phylum. Dark blue boxes highlight two species (humans and zebrafish). (C) Amino acid alignment between human and zebrafish cenexin C-terminal PLK1 binding motif. The serine highlighted in magenta represents the known human cenexin-PLK1 site (S796) and potential zebrafish biding site (S829). Letters in the middle between two sequences represent identical amino acids, and the + sign represent an amino acid of functional identity. Representative confocal maximum projection of cenexin from expanded (ExM) human cells and from a zebrafish embryo cell shown. Scale bar 0.05 μm. (D-F) Representative cells from 512-cell zebrafish embryos under cenexin depletion conditions (Cenexin MO, D), or rescue conditions (cenexin MO plus mCh-cenexin or <t>mCh-cenexin-S796A,</t> magenta in E-F) fixed and immunostained for γ-tubulin (inverted grey, D-E) or pST (inverted grey, F). Insets (D’, E’, E”, F’, and F”) at 5x magnification, corresponding areas outlined (μm 2 ). Scale bar, 5μm. (G) Scatter plot depicting γ-tubulin area (μm 2 ) at mitotic centrosomes under control, cenexin MO, and rescue conditions (cenexin MO plus mCh-cenexin or mCh-cenexin-S796A). Mean (magenta) with 95% confidence intervals shown. One-way ANOVA with multiple comparisons to control cells, n.s. not significant, **p
    Mcherry Cenexin S796a Plasmids, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs crispr cas9 skeleton vector
    <t>CRISPR/Cas9-based</t> deletion system in zebrafish. ( A ) Schematic illustration of the constructs of the CRISPR/Cas9-based vector, including the gfap promoter-driving Cas9, mCherry reporter, and U6-driving specific gRNA. ( B ) Strategy of the generation of the transgenic zebrafish lines. ( C ) F 1 transgenic fish generated by mosaic mCherry-positive fish outcrossed with wild-type fish. A subset of the transgenic offspring was identified by mCherry-positive astrocytes. Scale bar = 1 mm. ( D ) The colocalization of mCherry-labelled Cas9 and GFP-labelled endogenous Gfap in 60 hpf offspring from the cross of gfap WT and Tg ( gfap : GFP ) transgenic fish. Scale bars = 500 μm. ( E – J ) Co-localization of mCherry fluorescent signals (red) and endogenous Gfap detected by anti-Gfap antibody (FITC; green) in the brain and retina ( E – G ) or spinal cord ( H – J ) tissue. Scale bars = 200 μm. ( K ) Representative images of whole-mount in situ hybridization using an anti-sense RNA probe against Cas9 mRNA in 48 hpf embryos injected with Tol2 mRNA and gfap : Cas9-T2A-mCherry,U6 : gRNA ( null ) vector expressing Cas9 under the control of the gfap promoter. Cas9 expression pattern is governed by the tissue-specificity of gfap promoter (yellow arrowheads). Scale bars = 500 μm.
    Crispr Cas9 Skeleton Vector, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Identification of PTP1B (also called PTPN1) in the BioID and as interactor of RHBDL4. A , peptide overage of PTP1B in the BioID of RHBDL4. Peptides identified by MS are shown in blue. B , the substrate-trapping mutant D181A and the super-trapping mutant D181A/Y46F of mCherry-PTP1B were transiently expressed with or without RHBDL4mycFLAG. The cells were lysed in immunoprecipitation ( IP )–compatible conditions and captured with anti-FLAG immunomagnetic beads. Co-immunoprecipitates and whole cell lysates ( Input ) were probed with anti-mCherry, or RHBDL4 antibody. C , HEK293 cells expressing TMEM115-mycFLAG or RHBDL4mycFLAG were lysed and bound with anti-FLAG beads ± 4 m m Na 3 VO 4 . The resulting co-immunoprecipitates were probed with anti-FLAG-HRP or anti-mCherry antibody.

    Journal: The Journal of Biological Chemistry

    Article Title: Spatial proteomics reveal that the protein phosphatase PTP1B interacts with and may modify tyrosine phosphorylation of the rhomboid protease RHBDL4

    doi: 10.1074/jbc.RA118.007074

    Figure Lengend Snippet: Identification of PTP1B (also called PTPN1) in the BioID and as interactor of RHBDL4. A , peptide overage of PTP1B in the BioID of RHBDL4. Peptides identified by MS are shown in blue. B , the substrate-trapping mutant D181A and the super-trapping mutant D181A/Y46F of mCherry-PTP1B were transiently expressed with or without RHBDL4mycFLAG. The cells were lysed in immunoprecipitation ( IP )–compatible conditions and captured with anti-FLAG immunomagnetic beads. Co-immunoprecipitates and whole cell lysates ( Input ) were probed with anti-mCherry, or RHBDL4 antibody. C , HEK293 cells expressing TMEM115-mycFLAG or RHBDL4mycFLAG were lysed and bound with anti-FLAG beads ± 4 m m Na 3 VO 4 . The resulting co-immunoprecipitates were probed with anti-FLAG-HRP or anti-mCherry antibody.

    Article Snippet: The PCR product was cloned into SgfI/XhoI–excised RC210708 plasmid, by Gibson assembly (NEB). mCherry-PTP1B D181A was obtained from Addgene (40270). mCherry-PTP1B WT was generated by reverting the Ala181 codon back to aspartate, whereas super-trapping mutant D181A/Y46F was generated by site-directed mutagenesis of Tyr46 to Phe on the D181A mutant (Addgene, 40270).

    Techniques: Mutagenesis, Immunoprecipitation, Expressing

    Phenotypic and genetic analysis of G 1 offspring. ( a ) Freshly hatched G 1 offspring, one mCherry-negative chicken in the middle. ( b ) Distribution of mCherry-positive cells in the peripheral blood. FACS histogram of mCherry signal in red blood cells (blue) and white blood cells (red). ( c ) mCherry positivity visible in the beak and featherless skin of the mCherry-positive rooster (right) in the daylight. The mCherry-negative wt rooster shown as a control (left). ( d ) Photostability of mCherry shown as mCherry signal in adult feather. ( e ) The 195 bp PCR product of the CB-specific tpn allele amplified in DNA of G 1 offspring. The results of animals Nos 8 to 14 (from left to right) are shown, the non-CB chicken No. 11 is in the middle. Descendants of G 1 roosters Nos 769 and 795 are indicated. ( f ) Genomic relationship of inbred individual CB145 with itself, inbred individual CB151, the primordial germ cell line, the mCherry+ chicken Robin and a mCherry- chicken. The genomic relationship was calculated based on about 1.6 million maximum quality SNP detected in 78 chickens.

    Journal: Scientific Reports

    Article Title: Male fertility restored by transplanting primordial germ cells into testes: a new way towards efficient transgenesis in chicken

    doi: 10.1038/s41598-017-14475-w

    Figure Lengend Snippet: Phenotypic and genetic analysis of G 1 offspring. ( a ) Freshly hatched G 1 offspring, one mCherry-negative chicken in the middle. ( b ) Distribution of mCherry-positive cells in the peripheral blood. FACS histogram of mCherry signal in red blood cells (blue) and white blood cells (red). ( c ) mCherry positivity visible in the beak and featherless skin of the mCherry-positive rooster (right) in the daylight. The mCherry-negative wt rooster shown as a control (left). ( d ) Photostability of mCherry shown as mCherry signal in adult feather. ( e ) The 195 bp PCR product of the CB-specific tpn allele amplified in DNA of G 1 offspring. The results of animals Nos 8 to 14 (from left to right) are shown, the non-CB chicken No. 11 is in the middle. Descendants of G 1 roosters Nos 769 and 795 are indicated. ( f ) Genomic relationship of inbred individual CB145 with itself, inbred individual CB151, the primordial germ cell line, the mCherry+ chicken Robin and a mCherry- chicken. The genomic relationship was calculated based on about 1.6 million maximum quality SNP detected in 78 chickens.

    Article Snippet: An eGFP plasmid that was used before to generate transgenic chickens was used to swap the eGFP with the mCherry gene by Gibson Assembly (New England Biolabs) using the following oligos: fw: 5′-TATCGCATGCCTGCGATGGTGAGCAAGGGCGAGG-3′, rev: 5′-GCATGGACGAGCTGTACAAGTAGAACTTGTTTATTGCAGCT-3′.

    Techniques: FACS, Polymerase Chain Reaction, Amplification

    Analysis of modified PGCs and sperm of recipient rooster. ( a ) CB PGCs were transfected with a mCherry expression plasmid consisting of mCherry under a chicken β-actin promoter and a puromycin resistance gene under a CAG promoter flanked by HS4 insulators. ( b ) Succesful selection and expression of mCherry was confirmed by fluorescence microscopy and ( c ) flow cytometry. ( d ) The DNA junction between the mCherry vector and adjacent chicken genomic sequence detected in the whole-genome sequence of parental PGC clone. The vector-derived attB sequence is in bold and the repetitive motifs in the chicken genomic DNA are underlined. ( e ) mCherry positivity in the spermiogenic epithelium of G 0 recipient rooster after orthotopic transplantation of mCherry-positive CB PGCs. ( f ) The mCherry reporter gene was detected by PCR in mCherry+ PGCs and semen samples of two recipient roosters.

    Journal: Scientific Reports

    Article Title: Male fertility restored by transplanting primordial germ cells into testes: a new way towards efficient transgenesis in chicken

    doi: 10.1038/s41598-017-14475-w

    Figure Lengend Snippet: Analysis of modified PGCs and sperm of recipient rooster. ( a ) CB PGCs were transfected with a mCherry expression plasmid consisting of mCherry under a chicken β-actin promoter and a puromycin resistance gene under a CAG promoter flanked by HS4 insulators. ( b ) Succesful selection and expression of mCherry was confirmed by fluorescence microscopy and ( c ) flow cytometry. ( d ) The DNA junction between the mCherry vector and adjacent chicken genomic sequence detected in the whole-genome sequence of parental PGC clone. The vector-derived attB sequence is in bold and the repetitive motifs in the chicken genomic DNA are underlined. ( e ) mCherry positivity in the spermiogenic epithelium of G 0 recipient rooster after orthotopic transplantation of mCherry-positive CB PGCs. ( f ) The mCherry reporter gene was detected by PCR in mCherry+ PGCs and semen samples of two recipient roosters.

    Article Snippet: An eGFP plasmid that was used before to generate transgenic chickens was used to swap the eGFP with the mCherry gene by Gibson Assembly (New England Biolabs) using the following oligos: fw: 5′-TATCGCATGCCTGCGATGGTGAGCAAGGGCGAGG-3′, rev: 5′-GCATGGACGAGCTGTACAAGTAGAACTTGTTTATTGCAGCT-3′.

    Techniques: Modification, Transfection, Expressing, Plasmid Preparation, Selection, Fluorescence, Microscopy, Flow Cytometry, Sequencing, Pyrolysis Gas Chromatography, Derivative Assay, Transplantation Assay, Polymerase Chain Reaction

    ( a ) mCherry-positive embryo in the middle of incubation. ( b – e ) mCherry positivity in embryo organs and tissues. From left to right: ( b ) liver, ( c ) spleen, ( d ) skeletal muscle, ( e ) heart.

    Journal: Scientific Reports

    Article Title: Male fertility restored by transplanting primordial germ cells into testes: a new way towards efficient transgenesis in chicken

    doi: 10.1038/s41598-017-14475-w

    Figure Lengend Snippet: ( a ) mCherry-positive embryo in the middle of incubation. ( b – e ) mCherry positivity in embryo organs and tissues. From left to right: ( b ) liver, ( c ) spleen, ( d ) skeletal muscle, ( e ) heart.

    Article Snippet: An eGFP plasmid that was used before to generate transgenic chickens was used to swap the eGFP with the mCherry gene by Gibson Assembly (New England Biolabs) using the following oligos: fw: 5′-TATCGCATGCCTGCGATGGTGAGCAAGGGCGAGG-3′, rev: 5′-GCATGGACGAGCTGTACAAGTAGAACTTGTTTATTGCAGCT-3′.

    Techniques: Incubation

    Cenexin phosphorylation at its conserved C-terminal PLK1 binding site is required for maintenance of PCM in vivo . (A) Schematic representation of human hODF2 and cenexin (Cnxn, ODF2 isoform 9). The blue and magenta boxes highlight the N- and C-terminal extensions unique to cenexin. (B) Phylogenetic tree of the evolution of cenexin and its N-terminus and C-terminus in relation to Cep192 and centrin across different animal phyla. Cyan and orange boxes indicate whether cenexin and centrosome components (Cep192 and centrin) are detected or not detected within representative species of each phylum. Dark blue boxes highlight two species (humans and zebrafish). (C) Amino acid alignment between human and zebrafish cenexin C-terminal PLK1 binding motif. The serine highlighted in magenta represents the known human cenexin-PLK1 site (S796) and potential zebrafish biding site (S829). Letters in the middle between two sequences represent identical amino acids, and the + sign represent an amino acid of functional identity. Representative confocal maximum projection of cenexin from expanded (ExM) human cells and from a zebrafish embryo cell shown. Scale bar 0.05 μm. (D-F) Representative cells from 512-cell zebrafish embryos under cenexin depletion conditions (Cenexin MO, D), or rescue conditions (cenexin MO plus mCh-cenexin or mCh-cenexin-S796A, magenta in E-F) fixed and immunostained for γ-tubulin (inverted grey, D-E) or pST (inverted grey, F). Insets (D’, E’, E”, F’, and F”) at 5x magnification, corresponding areas outlined (μm 2 ). Scale bar, 5μm. (G) Scatter plot depicting γ-tubulin area (μm 2 ) at mitotic centrosomes under control, cenexin MO, and rescue conditions (cenexin MO plus mCh-cenexin or mCh-cenexin-S796A). Mean (magenta) with 95% confidence intervals shown. One-way ANOVA with multiple comparisons to control cells, n.s. not significant, **p

    Journal: bioRxiv

    Article Title: Pericentriolar matrix integrity relies on cenexin and Polo-Like Kinase (PLK)1

    doi: 10.1101/2022.01.09.475500

    Figure Lengend Snippet: Cenexin phosphorylation at its conserved C-terminal PLK1 binding site is required for maintenance of PCM in vivo . (A) Schematic representation of human hODF2 and cenexin (Cnxn, ODF2 isoform 9). The blue and magenta boxes highlight the N- and C-terminal extensions unique to cenexin. (B) Phylogenetic tree of the evolution of cenexin and its N-terminus and C-terminus in relation to Cep192 and centrin across different animal phyla. Cyan and orange boxes indicate whether cenexin and centrosome components (Cep192 and centrin) are detected or not detected within representative species of each phylum. Dark blue boxes highlight two species (humans and zebrafish). (C) Amino acid alignment between human and zebrafish cenexin C-terminal PLK1 binding motif. The serine highlighted in magenta represents the known human cenexin-PLK1 site (S796) and potential zebrafish biding site (S829). Letters in the middle between two sequences represent identical amino acids, and the + sign represent an amino acid of functional identity. Representative confocal maximum projection of cenexin from expanded (ExM) human cells and from a zebrafish embryo cell shown. Scale bar 0.05 μm. (D-F) Representative cells from 512-cell zebrafish embryos under cenexin depletion conditions (Cenexin MO, D), or rescue conditions (cenexin MO plus mCh-cenexin or mCh-cenexin-S796A, magenta in E-F) fixed and immunostained for γ-tubulin (inverted grey, D-E) or pST (inverted grey, F). Insets (D’, E’, E”, F’, and F”) at 5x magnification, corresponding areas outlined (μm 2 ). Scale bar, 5μm. (G) Scatter plot depicting γ-tubulin area (μm 2 ) at mitotic centrosomes under control, cenexin MO, and rescue conditions (cenexin MO plus mCh-cenexin or mCh-cenexin-S796A). Mean (magenta) with 95% confidence intervals shown. One-way ANOVA with multiple comparisons to control cells, n.s. not significant, **p

    Article Snippet: Plasmid Constructs and mRNA Gibson cloning methods were used to generate mCherry-cenexin-WT and mCherry-cenexin-S796A plasmids (NEBuilder HiFi DNA assembly kit), then purified using DNA maxi-prep kit (Bio Basic; 9K-006-0023). mRNA was generated from plasmids using mMESSAGE mMACHINE™SP6 transcription kit (Thermo Fisher Scientific; AM1340).

    Techniques: Binding Assay, In Vivo, Functional Assay

    CRISPR/Cas9-based deletion system in zebrafish. ( A ) Schematic illustration of the constructs of the CRISPR/Cas9-based vector, including the gfap promoter-driving Cas9, mCherry reporter, and U6-driving specific gRNA. ( B ) Strategy of the generation of the transgenic zebrafish lines. ( C ) F 1 transgenic fish generated by mosaic mCherry-positive fish outcrossed with wild-type fish. A subset of the transgenic offspring was identified by mCherry-positive astrocytes. Scale bar = 1 mm. ( D ) The colocalization of mCherry-labelled Cas9 and GFP-labelled endogenous Gfap in 60 hpf offspring from the cross of gfap WT and Tg ( gfap : GFP ) transgenic fish. Scale bars = 500 μm. ( E – J ) Co-localization of mCherry fluorescent signals (red) and endogenous Gfap detected by anti-Gfap antibody (FITC; green) in the brain and retina ( E – G ) or spinal cord ( H – J ) tissue. Scale bars = 200 μm. ( K ) Representative images of whole-mount in situ hybridization using an anti-sense RNA probe against Cas9 mRNA in 48 hpf embryos injected with Tol2 mRNA and gfap : Cas9-T2A-mCherry,U6 : gRNA ( null ) vector expressing Cas9 under the control of the gfap promoter. Cas9 expression pattern is governed by the tissue-specificity of gfap promoter (yellow arrowheads). Scale bars = 500 μm.

    Journal: Brain

    Article Title: Stepwise crosstalk between aberrant Nf1, Tp53 and Rb signalling pathways induces gliomagenesis in zebrafish

    doi: 10.1093/brain/awaa404

    Figure Lengend Snippet: CRISPR/Cas9-based deletion system in zebrafish. ( A ) Schematic illustration of the constructs of the CRISPR/Cas9-based vector, including the gfap promoter-driving Cas9, mCherry reporter, and U6-driving specific gRNA. ( B ) Strategy of the generation of the transgenic zebrafish lines. ( C ) F 1 transgenic fish generated by mosaic mCherry-positive fish outcrossed with wild-type fish. A subset of the transgenic offspring was identified by mCherry-positive astrocytes. Scale bar = 1 mm. ( D ) The colocalization of mCherry-labelled Cas9 and GFP-labelled endogenous Gfap in 60 hpf offspring from the cross of gfap WT and Tg ( gfap : GFP ) transgenic fish. Scale bars = 500 μm. ( E – J ) Co-localization of mCherry fluorescent signals (red) and endogenous Gfap detected by anti-Gfap antibody (FITC; green) in the brain and retina ( E – G ) or spinal cord ( H – J ) tissue. Scale bars = 200 μm. ( K ) Representative images of whole-mount in situ hybridization using an anti-sense RNA probe against Cas9 mRNA in 48 hpf embryos injected with Tol2 mRNA and gfap : Cas9-T2A-mCherry,U6 : gRNA ( null ) vector expressing Cas9 under the control of the gfap promoter. Cas9 expression pattern is governed by the tissue-specificity of gfap promoter (yellow arrowheads). Scale bars = 500 μm.

    Article Snippet: DNA construction and microinjection We cloned the zebrafish U6-3 promoter ( ) from the AB strain, followed by an NheI site, into the CRISPR/Cas9 skeleton vector (Cas9-T2A-mCherry ) from the Tol2 kit ( ) using ClaI and KpnI endonucleases (New England Biolabs).

    Techniques: CRISPR, Construct, Plasmid Preparation, Transgenic Assay, Fluorescence In Situ Hybridization, Generated, In Situ Hybridization, Injection, Expressing