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bsmbiigested lenticrisprv2 backbone  (Addgene inc)


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    Structured Review

    Addgene inc bsmbiigested lenticrisprv2 backbone
    Bsmbiigested Lenticrisprv2 Backbone, supplied by Addgene inc, used in various techniques. Bioz Stars score: 98/100, based on 527 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/lenticrisprv2+plasmid+backbone/lentiCRISPRv2+puro+(Plasmid+%2398290)/pm41903118-265-22-25
    Average 98 stars, based on 527 article reviews
    bsmbiigested lenticrisprv2 backbone - by Bioz Stars, 2026-11
    98/100 stars

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    Related Articles

    Plasmid Preparation:

    Article Title: The RNA sensor MDA5 detects SARS-CoV-2 infection
    Article Snippet: 48 h after transduction, cells were selected by addition of media containing 5 μg/ml puromycin (Gibco). .. The lentiCRISPRv2 plasmid backbone (Addgene #52,961) was used for generation of bulk knockout cell populations . .. The lentiCRISPRv2 plasmid backbone (Addgene #52,961) was used for generation of bulk knockout cell populations .

    Article Title: Epigenomes of Human Hearts Reveal New Genetic Variants Relevant for Cardiac Disease and Phenotype
    Article Snippet: .. The bioinformatic tool at http://crispor.tefor.net/ was used. sgRNA constructs were cloned into the Esp3I -digested LentiCRISPRv2 plasmid backbone (Addgene #52961) using T4 DNA ligase (catalogue #: M0202, New England Biolabs, NEB), according to manufacturer’s instructions. .. Vectors were transformed into RapidTrans TM TAM1 Competent E. coli (catalogue #: 11096, Active Motif).

    Article Title: The ETS transcription factor ETV6 constrains the transcriptional activity of EWS–FLI to promote Ewing sarcoma
    Article Snippet: RD cells were cultured in RPMI-1640 supplemented with 10% FBS. .. The lentiCRISPRv2 plasmid backbone (Addgene, 52961) encoding the Cas9 nuclease was digested with the restriction endonuclease BsmbI (Esp3I) (Thermo Fisher Scientific, FERFD0454) and gel extracted (Qiagen, 28704). ..

    Article Title: The RNA sensor MDA5 detects SARS-CoV-2 infection
    Article Snippet: 48 hours after transduction, cells were selected by addition of media containing 5 μg/ml puromycin (Gibco). .. The lentiCRISPRv2 plasmid backbone (Addgene #52961) was used for generation of bulk knockout cell populations . .. The lentiCRISPRv2 plasmid backbone (Addgene #52961) was used for generation of bulk knockout cell populations .

    Article Title: Epigenomes of Human Hearts Reveal New Genetic Variants Relevant for Cardiac Disease and Phenotype
    Article Snippet: .. The bioinformatic tool at D ow nloaded from http://ahajournals.org by on June 13, 2020 DOI: 10.1161/CIRCRESAHA.120.317254 5 http://crispor.tefor.net/ was used. sgRNA constructs were cloned into the Esp3I-digested LentiCRISPRv2 plasmid backbone (Addgene #52961) using T4 DNA ligase (catalogue #: M0202, New England Biolabs, NEB), according to manufacturer’s instructions. .. Vectors were transformed into RapidTransTM TAM1 Competent E. coli (catalogue #: 11096, Active Motif).

    Article Title: MYC-induced oncogenesis is dependent on acidic patches within its N-terminal intrinsically disordered domain
    Article Snippet: .. TRRAP CRISPR sgRNA guides sgRNA1: 5’-CAGCATTCCATCATTCCGA-3’, sgRNA2: 5’- CCACTGGGGATCGTTCAGTG-3’, sgRNA3: 5’- CTTGATCCGCCACTATACGA-3’, and sgRNA4: 5’- TGGTGTCAAGACAATCACGT-3’ from the Brunello Knock-Out library ( ) were cloned into a lentiCRISPRV2 plasmid backbone (Addgene # 244694). ..

    Knock-Out:

    Article Title: The RNA sensor MDA5 detects SARS-CoV-2 infection
    Article Snippet: 48 h after transduction, cells were selected by addition of media containing 5 μg/ml puromycin (Gibco). .. The lentiCRISPRv2 plasmid backbone (Addgene #52,961) was used for generation of bulk knockout cell populations . .. The lentiCRISPRv2 plasmid backbone (Addgene #52,961) was used for generation of bulk knockout cell populations .

    Article Title: The RNA sensor MDA5 detects SARS-CoV-2 infection
    Article Snippet: 48 hours after transduction, cells were selected by addition of media containing 5 μg/ml puromycin (Gibco). .. The lentiCRISPRv2 plasmid backbone (Addgene #52961) was used for generation of bulk knockout cell populations . .. The lentiCRISPRv2 plasmid backbone (Addgene #52961) was used for generation of bulk knockout cell populations .

    Article Title: MYC-induced oncogenesis is dependent on acidic patches within its N-terminal intrinsically disordered domain
    Article Snippet: .. TRRAP CRISPR sgRNA guides sgRNA1: 5’-CAGCATTCCATCATTCCGA-3’, sgRNA2: 5’- CCACTGGGGATCGTTCAGTG-3’, sgRNA3: 5’- CTTGATCCGCCACTATACGA-3’, and sgRNA4: 5’- TGGTGTCAAGACAATCACGT-3’ from the Brunello Knock-Out library ( ) were cloned into a lentiCRISPRV2 plasmid backbone (Addgene # 244694). ..

    Construct:

    Article Title: Epigenomes of Human Hearts Reveal New Genetic Variants Relevant for Cardiac Disease and Phenotype
    Article Snippet: .. The bioinformatic tool at http://crispor.tefor.net/ was used. sgRNA constructs were cloned into the Esp3I -digested LentiCRISPRv2 plasmid backbone (Addgene #52961) using T4 DNA ligase (catalogue #: M0202, New England Biolabs, NEB), according to manufacturer’s instructions. .. Vectors were transformed into RapidTrans TM TAM1 Competent E. coli (catalogue #: 11096, Active Motif).

    Article Title: Epigenomes of Human Hearts Reveal New Genetic Variants Relevant for Cardiac Disease and Phenotype
    Article Snippet: .. The bioinformatic tool at D ow nloaded from http://ahajournals.org by on June 13, 2020 DOI: 10.1161/CIRCRESAHA.120.317254 5 http://crispor.tefor.net/ was used. sgRNA constructs were cloned into the Esp3I-digested LentiCRISPRv2 plasmid backbone (Addgene #52961) using T4 DNA ligase (catalogue #: M0202, New England Biolabs, NEB), according to manufacturer’s instructions. .. Vectors were transformed into RapidTransTM TAM1 Competent E. coli (catalogue #: 11096, Active Motif).

    Clone Assay:

    Article Title: Epigenomes of Human Hearts Reveal New Genetic Variants Relevant for Cardiac Disease and Phenotype
    Article Snippet: .. The bioinformatic tool at http://crispor.tefor.net/ was used. sgRNA constructs were cloned into the Esp3I -digested LentiCRISPRv2 plasmid backbone (Addgene #52961) using T4 DNA ligase (catalogue #: M0202, New England Biolabs, NEB), according to manufacturer’s instructions. .. Vectors were transformed into RapidTrans TM TAM1 Competent E. coli (catalogue #: 11096, Active Motif).

    Article Title: Epigenomes of Human Hearts Reveal New Genetic Variants Relevant for Cardiac Disease and Phenotype
    Article Snippet: .. The bioinformatic tool at D ow nloaded from http://ahajournals.org by on June 13, 2020 DOI: 10.1161/CIRCRESAHA.120.317254 5 http://crispor.tefor.net/ was used. sgRNA constructs were cloned into the Esp3I-digested LentiCRISPRv2 plasmid backbone (Addgene #52961) using T4 DNA ligase (catalogue #: M0202, New England Biolabs, NEB), according to manufacturer’s instructions. .. Vectors were transformed into RapidTransTM TAM1 Competent E. coli (catalogue #: 11096, Active Motif).

    Article Title: MYC-induced oncogenesis is dependent on acidic patches within its N-terminal intrinsically disordered domain
    Article Snippet: .. TRRAP CRISPR sgRNA guides sgRNA1: 5’-CAGCATTCCATCATTCCGA-3’, sgRNA2: 5’- CCACTGGGGATCGTTCAGTG-3’, sgRNA3: 5’- CTTGATCCGCCACTATACGA-3’, and sgRNA4: 5’- TGGTGTCAAGACAATCACGT-3’ from the Brunello Knock-Out library ( ) were cloned into a lentiCRISPRV2 plasmid backbone (Addgene # 244694). ..

    CRISPR:

    Article Title: MYC-induced oncogenesis is dependent on acidic patches within its N-terminal intrinsically disordered domain
    Article Snippet: .. TRRAP CRISPR sgRNA guides sgRNA1: 5’-CAGCATTCCATCATTCCGA-3’, sgRNA2: 5’- CCACTGGGGATCGTTCAGTG-3’, sgRNA3: 5’- CTTGATCCGCCACTATACGA-3’, and sgRNA4: 5’- TGGTGTCAAGACAATCACGT-3’ from the Brunello Knock-Out library ( ) were cloned into a lentiCRISPRV2 plasmid backbone (Addgene # 244694). ..



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    MYC regulates ASS1 expression in the context of BRAF/MEK inhibition . ( a and b ) Melanoma cells were treated with Dabrafenib/Trametinib (D/T) for 5 days. Total mRNAs were isolated and subjected to qPCR to detect the mRNA expression of ASS1 and MYC ( a ). The same treatment was performed, protein lysates were collected, and ASS1 and MYC protein expression were detected by western blot ( b ). β-ACTIN was used as a loading control. ( c-f ) Non-targeting sgRNA (sgNT) or sgRNAs targeting MYC (sgMYC) were designed, cloned <t>to</t> <t>lentiCRISPRv2-blasticidin</t> vector, used to package lentivirus, and infect target melanoma cells. After blasticidin selection, total mRNAs from survived cells were isolated and subjected to qPCR to quantify mRNA expression of ASS1 ( c ), while protein lysates were collected and used for western blot to detect protein expression of ASS1 and MYC ( d ). sgAss1 was used as a positive control. β-ACTIN was used as a loading control. Meanwhile, blasticidin resistant cells were seeded onto 6-well plates, treated with PBS or ADI-PEG20 for 10 days, and crystal violet was used to stain colonies formed. Representative images were shown ( e ) and area percentage of stained colonies were quantified ( f ).
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    Image Search Results


    MYC regulates ASS1 expression in the context of BRAF/MEK inhibition . ( a and b ) Melanoma cells were treated with Dabrafenib/Trametinib (D/T) for 5 days. Total mRNAs were isolated and subjected to qPCR to detect the mRNA expression of ASS1 and MYC ( a ). The same treatment was performed, protein lysates were collected, and ASS1 and MYC protein expression were detected by western blot ( b ). β-ACTIN was used as a loading control. ( c-f ) Non-targeting sgRNA (sgNT) or sgRNAs targeting MYC (sgMYC) were designed, cloned to lentiCRISPRv2-blasticidin vector, used to package lentivirus, and infect target melanoma cells. After blasticidin selection, total mRNAs from survived cells were isolated and subjected to qPCR to quantify mRNA expression of ASS1 ( c ), while protein lysates were collected and used for western blot to detect protein expression of ASS1 and MYC ( d ). sgAss1 was used as a positive control. β-ACTIN was used as a loading control. Meanwhile, blasticidin resistant cells were seeded onto 6-well plates, treated with PBS or ADI-PEG20 for 10 days, and crystal violet was used to stain colonies formed. Representative images were shown ( e ) and area percentage of stained colonies were quantified ( f ).

    Journal: bioRxiv

    Article Title: MYC-ATF4-ASS1 axis governs intracellular arginine synthesis and dictates the immune microenvironment in melanoma

    doi: 10.64898/2026.02.27.707779

    Figure Lengend Snippet: MYC regulates ASS1 expression in the context of BRAF/MEK inhibition . ( a and b ) Melanoma cells were treated with Dabrafenib/Trametinib (D/T) for 5 days. Total mRNAs were isolated and subjected to qPCR to detect the mRNA expression of ASS1 and MYC ( a ). The same treatment was performed, protein lysates were collected, and ASS1 and MYC protein expression were detected by western blot ( b ). β-ACTIN was used as a loading control. ( c-f ) Non-targeting sgRNA (sgNT) or sgRNAs targeting MYC (sgMYC) were designed, cloned to lentiCRISPRv2-blasticidin vector, used to package lentivirus, and infect target melanoma cells. After blasticidin selection, total mRNAs from survived cells were isolated and subjected to qPCR to quantify mRNA expression of ASS1 ( c ), while protein lysates were collected and used for western blot to detect protein expression of ASS1 and MYC ( d ). sgAss1 was used as a positive control. β-ACTIN was used as a loading control. Meanwhile, blasticidin resistant cells were seeded onto 6-well plates, treated with PBS or ADI-PEG20 for 10 days, and crystal violet was used to stain colonies formed. Representative images were shown ( e ) and area percentage of stained colonies were quantified ( f ).

    Article Snippet: Briefly, non-targeting or gene-specific targeting oligos were annealed and ligated into a lentiCRISPRv2-blasticidin backbone (Addgene, 98293) cut with BsmBI-v2 (NEB, R0739S).

    Techniques: Expressing, Inhibition, Isolation, Western Blot, Control, Clone Assay, Plasmid Preparation, Selection, Positive Control, Staining

    ATF4 correlates with and regulates ASS1 expression . ( a ) WM4380-2 cells were treated with DMSO, D/T, ADI or D/T+ADI as shown in . Total mRNA were isolated and subjected to bulk RNA-seq. Normalized reads of ATF4 were presented. ( b ) Correlative analysis of ATF4 and ASS1 was performed using our bulk RNA-seq data as indicated in . R and P value were shown. ( c ) Correlative analysis of ATF4 and ASS1 was performed exploiting public dataset of spatical single cell RNA-seq. R and P valude were provided. ( d-f ) Non-targeting sgRNA (sgNT) or sgRNAs targeting ATF4 (sgATF4) were designed, cloned to lentiCRISPRv2-blasticidin vector, used to package lentivirus, and infect target melanoma cells. After blasticidin selection, protein lysates were collected and used for western blot to detect protein expression of ASS1 and ATF4 ( d ). β-ACTIN was used as a loading control. Meanwhile, blasticidin resistant cells were seeded onto 6-well plates, treated with PBS or ADI-PEG20 for 10 days, and crystal violet was used to stain colonies formed. Representative images were shown ( e ) and area percentage of stained colonies were quantified ( f ).

    Journal: bioRxiv

    Article Title: MYC-ATF4-ASS1 axis governs intracellular arginine synthesis and dictates the immune microenvironment in melanoma

    doi: 10.64898/2026.02.27.707779

    Figure Lengend Snippet: ATF4 correlates with and regulates ASS1 expression . ( a ) WM4380-2 cells were treated with DMSO, D/T, ADI or D/T+ADI as shown in . Total mRNA were isolated and subjected to bulk RNA-seq. Normalized reads of ATF4 were presented. ( b ) Correlative analysis of ATF4 and ASS1 was performed using our bulk RNA-seq data as indicated in . R and P value were shown. ( c ) Correlative analysis of ATF4 and ASS1 was performed exploiting public dataset of spatical single cell RNA-seq. R and P valude were provided. ( d-f ) Non-targeting sgRNA (sgNT) or sgRNAs targeting ATF4 (sgATF4) were designed, cloned to lentiCRISPRv2-blasticidin vector, used to package lentivirus, and infect target melanoma cells. After blasticidin selection, protein lysates were collected and used for western blot to detect protein expression of ASS1 and ATF4 ( d ). β-ACTIN was used as a loading control. Meanwhile, blasticidin resistant cells were seeded onto 6-well plates, treated with PBS or ADI-PEG20 for 10 days, and crystal violet was used to stain colonies formed. Representative images were shown ( e ) and area percentage of stained colonies were quantified ( f ).

    Article Snippet: Briefly, non-targeting or gene-specific targeting oligos were annealed and ligated into a lentiCRISPRv2-blasticidin backbone (Addgene, 98293) cut with BsmBI-v2 (NEB, R0739S).

    Techniques: Expressing, Isolation, RNA Sequencing, Single Cell, Clone Assay, Plasmid Preparation, Selection, Western Blot, Control, Staining