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guide rna ngrna backbones  (Addgene inc)


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    Structured Review

    Addgene inc guide rna ngrna backbones
    Guide Rna Ngrna Backbones, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 103 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/guide+rna+plasmid+backbone/BPK1520+(Plasmid+%2365777)/pmc12825063-121-7-13
    Average 94 stars, based on 103 article reviews
    guide rna ngrna backbones - by Bioz Stars, 2026-09
    94/100 stars

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    Related Articles

    Plasmid Preparation:

    Article Title: Configuration of adaptable template RNA architectures to unfold the editable space of a nuclease prime editor.
    Article Snippet: For fragments < 1 kb, the Phanta Max Master Mix (Vazyme) was employed. .. The guide RNA plasmid backbone was amplified from pGL3-U6sgRNA-EGFP (Addgene #107 721). .. The assembly of the various pegRNA or sgRNA cassettes was conducted by preparation of annealed primers, followed by a step of cassette assembly into the backbone vector aided by the MultiF Seamless Assembly Mix (ABclonal).

    Article Title: An upgraded nuclease prime editor platform enables high-efficiency singled or multiplexed knock-in/knockout of genes in mouse and sheep zygotes
    Article Snippet: During vector construction, the Phanta Flash Master Mix (Vazyme) was used for PCR amplification of products greater than 1 kb, and the Phanta Max Master Mix (Vazyme) was used for fragments less than 1 kb. .. The guide RNA plasmid backbone was amplified from pGL3- U6-sgRNA-EGFP (Addgene #107721). .. The pegRNA cassette or the sgRNA cassette were first assembled using primers and then cloned into the backbone vector using MultiF Seamless Assembly Mix (ABclonal).

    Article Title: Configuration of adaptable template RNA architectures to unfold the editable space of a nuclease prime editor
    Article Snippet: For fragments < 1 kb, the Phanta Max Master Mix (Vazyme) was employed. .. The guide RNA plasmid backbone was amplified from pGL3-U6-sgRNA-EGFP (Addgene #107 721). .. The assembly of the various pegRNA or sgRNA cassettes was conducted by preparation of annealed primers, followed by a step of cassette assembly into the backbone vector aided by the MultiF Seamless Assembly Mix (ABclonal).

    Amplification:

    Article Title: Configuration of adaptable template RNA architectures to unfold the editable space of a nuclease prime editor.
    Article Snippet: For fragments < 1 kb, the Phanta Max Master Mix (Vazyme) was employed. .. The guide RNA plasmid backbone was amplified from pGL3-U6sgRNA-EGFP (Addgene #107 721). .. The assembly of the various pegRNA or sgRNA cassettes was conducted by preparation of annealed primers, followed by a step of cassette assembly into the backbone vector aided by the MultiF Seamless Assembly Mix (ABclonal).

    Article Title: An upgraded nuclease prime editor platform enables high-efficiency singled or multiplexed knock-in/knockout of genes in mouse and sheep zygotes
    Article Snippet: During vector construction, the Phanta Flash Master Mix (Vazyme) was used for PCR amplification of products greater than 1 kb, and the Phanta Max Master Mix (Vazyme) was used for fragments less than 1 kb. .. The guide RNA plasmid backbone was amplified from pGL3- U6-sgRNA-EGFP (Addgene #107721). .. The pegRNA cassette or the sgRNA cassette were first assembled using primers and then cloned into the backbone vector using MultiF Seamless Assembly Mix (ABclonal).

    Article Title: Configuration of adaptable template RNA architectures to unfold the editable space of a nuclease prime editor
    Article Snippet: For fragments < 1 kb, the Phanta Max Master Mix (Vazyme) was employed. .. The guide RNA plasmid backbone was amplified from pGL3-U6-sgRNA-EGFP (Addgene #107 721). .. The assembly of the various pegRNA or sgRNA cassettes was conducted by preparation of annealed primers, followed by a step of cassette assembly into the backbone vector aided by the MultiF Seamless Assembly Mix (ABclonal).



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    Effects of dCas9-KM system on reporter gene expression in embryos of X. tropicalis . ( A ) Schematic of CRISPRi-related gRNAs targeting the regions of mCherry reporter gene in pmCherry-N1 plasmid. ( B ) Schematic illustration of the experimental setup used to analyze dCas9-KM capacity targeting exogenous reporter gene in X. tropicalis embryos. ( C ) The qPCR validation of mCherry expression in embryos co-injected with dCas9-KM system and indicated gRNAs at 48 dpi ( n = 4 per group). ( D and E ) Representative images ( D ) and quantification ( E ) of mCherry expression in embryos co-injected with dCas9-KM system and indicated <t>gRNA</t> at 48 hpi ( n = ~ 60 embryos from 3 independent experiments). ( F ) Schematic of gR-187 targeting mCherry reporter gene in pCMV-mCherry-EF1α-EGFP dual-reporter plasmid. ( G ) The qPCR validation of mCherry expression in embryos co-injected with dCas9-KM system and indicated gRNA at 48 dpi ( n = 5 per group). EGFP was used as the internal control. ( H and I ) Representative images ( H ) and quantification ( I ) of mCherry fluorescence in embryos co-injected with dCas9-KM system and indicate gRNA at 48 hpi ( n = 24 embryos from 3 independent experiments). All data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus control. Ns, no significant differences versus control
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    Effects of dCas9-KM system on reporter gene expression in embryos of X. tropicalis . ( A ) Schematic of CRISPRi-related gRNAs targeting the regions of mCherry reporter gene in pmCherry-N1 plasmid. ( B ) Schematic illustration of the experimental setup used to analyze dCas9-KM capacity targeting exogenous reporter gene in X. tropicalis embryos. ( C ) The qPCR validation of mCherry expression in embryos co-injected with dCas9-KM system and indicated gRNAs at 48 dpi ( n = 4 per group). ( D and E ) Representative images ( D ) and quantification ( E ) of mCherry expression in embryos co-injected with dCas9-KM system and indicated <t>gRNA</t> at 48 hpi ( n = ~ 60 embryos from 3 independent experiments). ( F ) Schematic of gR-187 targeting mCherry reporter gene in pCMV-mCherry-EF1α-EGFP dual-reporter plasmid. ( G ) The qPCR validation of mCherry expression in embryos co-injected with dCas9-KM system and indicated gRNA at 48 dpi ( n = 5 per group). EGFP was used as the internal control. ( H and I ) Representative images ( H ) and quantification ( I ) of mCherry fluorescence in embryos co-injected with dCas9-KM system and indicate gRNA at 48 hpi ( n = 24 embryos from 3 independent experiments). All data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus control. Ns, no significant differences versus control
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    Image Search Results


    Effects of dCas9-KM system on reporter gene expression in embryos of X. tropicalis . ( A ) Schematic of CRISPRi-related gRNAs targeting the regions of mCherry reporter gene in pmCherry-N1 plasmid. ( B ) Schematic illustration of the experimental setup used to analyze dCas9-KM capacity targeting exogenous reporter gene in X. tropicalis embryos. ( C ) The qPCR validation of mCherry expression in embryos co-injected with dCas9-KM system and indicated gRNAs at 48 dpi ( n = 4 per group). ( D and E ) Representative images ( D ) and quantification ( E ) of mCherry expression in embryos co-injected with dCas9-KM system and indicated gRNA at 48 hpi ( n = ~ 60 embryos from 3 independent experiments). ( F ) Schematic of gR-187 targeting mCherry reporter gene in pCMV-mCherry-EF1α-EGFP dual-reporter plasmid. ( G ) The qPCR validation of mCherry expression in embryos co-injected with dCas9-KM system and indicated gRNA at 48 dpi ( n = 5 per group). EGFP was used as the internal control. ( H and I ) Representative images ( H ) and quantification ( I ) of mCherry fluorescence in embryos co-injected with dCas9-KM system and indicate gRNA at 48 hpi ( n = 24 embryos from 3 independent experiments). All data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus control. Ns, no significant differences versus control

    Journal: Cell & Bioscience

    Article Title: CRISPR/Cas-mediated mRNA knockdown in the embryos of Xenopus tropicalis

    doi: 10.1186/s13578-025-01397-8

    Figure Lengend Snippet: Effects of dCas9-KM system on reporter gene expression in embryos of X. tropicalis . ( A ) Schematic of CRISPRi-related gRNAs targeting the regions of mCherry reporter gene in pmCherry-N1 plasmid. ( B ) Schematic illustration of the experimental setup used to analyze dCas9-KM capacity targeting exogenous reporter gene in X. tropicalis embryos. ( C ) The qPCR validation of mCherry expression in embryos co-injected with dCas9-KM system and indicated gRNAs at 48 dpi ( n = 4 per group). ( D and E ) Representative images ( D ) and quantification ( E ) of mCherry expression in embryos co-injected with dCas9-KM system and indicated gRNA at 48 hpi ( n = ~ 60 embryos from 3 independent experiments). ( F ) Schematic of gR-187 targeting mCherry reporter gene in pCMV-mCherry-EF1α-EGFP dual-reporter plasmid. ( G ) The qPCR validation of mCherry expression in embryos co-injected with dCas9-KM system and indicated gRNA at 48 dpi ( n = 5 per group). EGFP was used as the internal control. ( H and I ) Representative images ( H ) and quantification ( I ) of mCherry fluorescence in embryos co-injected with dCas9-KM system and indicate gRNA at 48 hpi ( n = 24 embryos from 3 independent experiments). All data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus control. Ns, no significant differences versus control

    Article Snippet: The human codon-optimized PspCas13b (pC0046, #103862) and RfxCas13d (pXR001, #109049) expressing plasmids as well as their guide RNA (gRNA) expression plasmids (pC0043, #103854 for PspCas13b; pXR003, #109053 for RfxCas13d) were purchased from Addgene.

    Techniques: Gene Expression, Plasmid Preparation, Biomarker Discovery, Expressing, Injection, Control, Fluorescence

    Effects of dCas9-KM system on tyrosinase expression in embryos of X. tropicalis. ( A ) Schematic illustration of dCas9-KM system-related gRNAs targeting the DNA locus of endogenous tyrosinase (Tyr) gene. Fertilized eggs were co-injected with dCas9-KM mRNAs and indicated gRNA sets, followed by embryo evaluation at 48 hpi as follows. ( B and C ) Representative images ( B ) and quantification ( C ) of Tyr production in embryos co-injected with dCas9-KM mRNA and indicated gRNAs at 48 hpi ( n = 26 ~ 30 embryos from 3 independent experiments). Scale bar = 250 μm. ( D ) The qPCR validation of Tyr expression in embryos co-injected with dCas9-KM mRNA and indicated gRNAs at 48 dpi ( n = 5 per group). ( E and F ) Representative images ( E , scale bar = 250 μm) and quantification ( F ) of embryos with different phenotypes were counted and compared with the total developed ones 48 h post co-injection of dCas9-KM system. ( G and H ) Representative images ( G ) and quantification ( H ) of embryos with different phenotypes were counted and compared with the total developed ones 72 h post co-injection of dCas9-KM system. Total embryos evaluated for each group (n) is shown above each column. All data are present as mean ± SEM. **** p < 0.0001 versus gNC group

    Journal: Cell & Bioscience

    Article Title: CRISPR/Cas-mediated mRNA knockdown in the embryos of Xenopus tropicalis

    doi: 10.1186/s13578-025-01397-8

    Figure Lengend Snippet: Effects of dCas9-KM system on tyrosinase expression in embryos of X. tropicalis. ( A ) Schematic illustration of dCas9-KM system-related gRNAs targeting the DNA locus of endogenous tyrosinase (Tyr) gene. Fertilized eggs were co-injected with dCas9-KM mRNAs and indicated gRNA sets, followed by embryo evaluation at 48 hpi as follows. ( B and C ) Representative images ( B ) and quantification ( C ) of Tyr production in embryos co-injected with dCas9-KM mRNA and indicated gRNAs at 48 hpi ( n = 26 ~ 30 embryos from 3 independent experiments). Scale bar = 250 μm. ( D ) The qPCR validation of Tyr expression in embryos co-injected with dCas9-KM mRNA and indicated gRNAs at 48 dpi ( n = 5 per group). ( E and F ) Representative images ( E , scale bar = 250 μm) and quantification ( F ) of embryos with different phenotypes were counted and compared with the total developed ones 48 h post co-injection of dCas9-KM system. ( G and H ) Representative images ( G ) and quantification ( H ) of embryos with different phenotypes were counted and compared with the total developed ones 72 h post co-injection of dCas9-KM system. Total embryos evaluated for each group (n) is shown above each column. All data are present as mean ± SEM. **** p < 0.0001 versus gNC group

    Article Snippet: The human codon-optimized PspCas13b (pC0046, #103862) and RfxCas13d (pXR001, #109049) expressing plasmids as well as their guide RNA (gRNA) expression plasmids (pC0043, #103854 for PspCas13b; pXR003, #109053 for RfxCas13d) were purchased from Addgene.

    Techniques: Expressing, Injection, Biomarker Discovery

    Effects of dCas9-KM system on pax6 expression in embryos of X. tropicalis . ( A ) Schematic illustration of dCas9-KM system-related gRNAs targeting the DNA locus of endogenous pax6 gene. Fertilized eggs were co-injected with dCas9-KM mRNAs and indicated gRNA sets, followed by embryo evaluation at 48 hpi as follows. ( B ) The qPCR validation of pax6 expression in the dCas9-KM-injected embryos with different gRNA sets ( n = 5 per group). ( C ) Representative images of embryos with smaller eyes in the experimental group injected with pax6 targeting gRNA sets. The top panel shows low magnification, and the lower panel shows high magnification. Asterisk denotes smaller eyes. ( D ) Embryos with different phenotypes were counted and compared with the total developed ones after injection. Total embryos evaluated for each group (n) is shown above each column. ( E ) Quantification of eye size in tadpoles treated with gRNA sets and controls ( n = 3 per group). ( F ) The qPCR validation of pax6 expression in the dCas9-KM-injected embryos with single gRNAs from set2 ( n = 5 per group). All data are present as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 versus gNC group. Ns, no significant differences versus gNC group

    Journal: Cell & Bioscience

    Article Title: CRISPR/Cas-mediated mRNA knockdown in the embryos of Xenopus tropicalis

    doi: 10.1186/s13578-025-01397-8

    Figure Lengend Snippet: Effects of dCas9-KM system on pax6 expression in embryos of X. tropicalis . ( A ) Schematic illustration of dCas9-KM system-related gRNAs targeting the DNA locus of endogenous pax6 gene. Fertilized eggs were co-injected with dCas9-KM mRNAs and indicated gRNA sets, followed by embryo evaluation at 48 hpi as follows. ( B ) The qPCR validation of pax6 expression in the dCas9-KM-injected embryos with different gRNA sets ( n = 5 per group). ( C ) Representative images of embryos with smaller eyes in the experimental group injected with pax6 targeting gRNA sets. The top panel shows low magnification, and the lower panel shows high magnification. Asterisk denotes smaller eyes. ( D ) Embryos with different phenotypes were counted and compared with the total developed ones after injection. Total embryos evaluated for each group (n) is shown above each column. ( E ) Quantification of eye size in tadpoles treated with gRNA sets and controls ( n = 3 per group). ( F ) The qPCR validation of pax6 expression in the dCas9-KM-injected embryos with single gRNAs from set2 ( n = 5 per group). All data are present as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 versus gNC group. Ns, no significant differences versus gNC group

    Article Snippet: The human codon-optimized PspCas13b (pC0046, #103862) and RfxCas13d (pXR001, #109049) expressing plasmids as well as their guide RNA (gRNA) expression plasmids (pC0043, #103854 for PspCas13b; pXR003, #109053 for RfxCas13d) were purchased from Addgene.

    Techniques: Expressing, Injection, Biomarker Discovery