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zim3krab domain  (Addgene inc)


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    Addgene inc zim3krab domain
    Zim3krab Domain, supplied by Addgene inc, used in various techniques. Bioz Stars score: 95/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/zim3krab+dcas9+domain/pLX303-ZIM3-KRAB-dCas9+(Plasmid+%23154472)/bio_rxiv__64898__2026__01__27__702006-319-7-11
    Average 95 stars, based on 32 article reviews
    zim3krab domain - by Bioz Stars, 2026-10
    95/100 stars

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    Related Articles

    Plasmid Preparation:

    Article Title: Systematic decoding of cis gene regulation defines context-dependent control of the multi-gene costimulatory receptor locus in human T cells.
    Article Snippet: Protospacers flanked by adaptor sequences were synthesized by Agilent Technologies, amplified by PCR and cloned into the pCRISPRia-v2 lentiviral vector (Addgene, 84832) using BstXI (NEB, R0113) and Blpl (NEB, R0585)97. .. The dCas9-ZIM3 plasmid was constructed using NEBuilder HiFi DNA Assembly Master Mix (NEB, E2621), the sequence for the ZIM3KRAB–dCas9 domain amplified from pLX303-ZIM3-KRAB-dCas9 (Addgene, 154472) by PCR with primers CM_oligo_1 and CM_oligo_2 (Supplementary Table 1) and the Lenti-SFFV-mCherry-dCas9-VP64 (ref. 46) (Addgene, 180263) lentiviral backbone digested with PmeI (NEB, R0560) and BamHI (NEB, R3136). .. For arrayed CRISPRi validation experiments, sgRNA sequences were ordered as Ultramers (IDT, 0.2 μM) with flanking adaptors, as in CM_oligo_3, cloned into the pCRISPRia-v2 lentiviral vector (Addgene, 84832) digested with Blpl (NEB, R0585) and BstXI (NEB, R0113) using NEBuilder HiFi DNA Assembly Master Mix (NEB, E2621) and transformed into STBL3 chemically competent cells (QB3 MacroLab).

    Construct:

    Article Title: Systematic decoding of cis gene regulation defines context-dependent control of the multi-gene costimulatory receptor locus in human T cells.
    Article Snippet: Protospacers flanked by adaptor sequences were synthesized by Agilent Technologies, amplified by PCR and cloned into the pCRISPRia-v2 lentiviral vector (Addgene, 84832) using BstXI (NEB, R0113) and Blpl (NEB, R0585)97. .. The dCas9-ZIM3 plasmid was constructed using NEBuilder HiFi DNA Assembly Master Mix (NEB, E2621), the sequence for the ZIM3KRAB–dCas9 domain amplified from pLX303-ZIM3-KRAB-dCas9 (Addgene, 154472) by PCR with primers CM_oligo_1 and CM_oligo_2 (Supplementary Table 1) and the Lenti-SFFV-mCherry-dCas9-VP64 (ref. 46) (Addgene, 180263) lentiviral backbone digested with PmeI (NEB, R0560) and BamHI (NEB, R3136). .. For arrayed CRISPRi validation experiments, sgRNA sequences were ordered as Ultramers (IDT, 0.2 μM) with flanking adaptors, as in CM_oligo_3, cloned into the pCRISPRia-v2 lentiviral vector (Addgene, 84832) digested with Blpl (NEB, R0585) and BstXI (NEB, R0113) using NEBuilder HiFi DNA Assembly Master Mix (NEB, E2621) and transformed into STBL3 chemically competent cells (QB3 MacroLab).

    Sequencing:

    Article Title: Systematic decoding of cis gene regulation defines context-dependent control of the multi-gene costimulatory receptor locus in human T cells.
    Article Snippet: Protospacers flanked by adaptor sequences were synthesized by Agilent Technologies, amplified by PCR and cloned into the pCRISPRia-v2 lentiviral vector (Addgene, 84832) using BstXI (NEB, R0113) and Blpl (NEB, R0585)97. .. The dCas9-ZIM3 plasmid was constructed using NEBuilder HiFi DNA Assembly Master Mix (NEB, E2621), the sequence for the ZIM3KRAB–dCas9 domain amplified from pLX303-ZIM3-KRAB-dCas9 (Addgene, 154472) by PCR with primers CM_oligo_1 and CM_oligo_2 (Supplementary Table 1) and the Lenti-SFFV-mCherry-dCas9-VP64 (ref. 46) (Addgene, 180263) lentiviral backbone digested with PmeI (NEB, R0560) and BamHI (NEB, R3136). .. For arrayed CRISPRi validation experiments, sgRNA sequences were ordered as Ultramers (IDT, 0.2 μM) with flanking adaptors, as in CM_oligo_3, cloned into the pCRISPRia-v2 lentiviral vector (Addgene, 84832) digested with Blpl (NEB, R0585) and BstXI (NEB, R0113) using NEBuilder HiFi DNA Assembly Master Mix (NEB, E2621) and transformed into STBL3 chemically competent cells (QB3 MacroLab).

    Amplification:

    Article Title: Systematic decoding of cis gene regulation defines context-dependent control of the multi-gene costimulatory receptor locus in human T cells.
    Article Snippet: Protospacers flanked by adaptor sequences were synthesized by Agilent Technologies, amplified by PCR and cloned into the pCRISPRia-v2 lentiviral vector (Addgene, 84832) using BstXI (NEB, R0113) and Blpl (NEB, R0585)97. .. The dCas9-ZIM3 plasmid was constructed using NEBuilder HiFi DNA Assembly Master Mix (NEB, E2621), the sequence for the ZIM3KRAB–dCas9 domain amplified from pLX303-ZIM3-KRAB-dCas9 (Addgene, 154472) by PCR with primers CM_oligo_1 and CM_oligo_2 (Supplementary Table 1) and the Lenti-SFFV-mCherry-dCas9-VP64 (ref. 46) (Addgene, 180263) lentiviral backbone digested with PmeI (NEB, R0560) and BamHI (NEB, R3136). .. For arrayed CRISPRi validation experiments, sgRNA sequences were ordered as Ultramers (IDT, 0.2 μM) with flanking adaptors, as in CM_oligo_3, cloned into the pCRISPRia-v2 lentiviral vector (Addgene, 84832) digested with Blpl (NEB, R0585) and BstXI (NEB, R0113) using NEBuilder HiFi DNA Assembly Master Mix (NEB, E2621) and transformed into STBL3 chemically competent cells (QB3 MacroLab).

    Polymerase Chain Reaction:

    Article Title: Systematic decoding of cis gene regulation defines context-dependent control of the multi-gene costimulatory receptor locus in human T cells.
    Article Snippet: Protospacers flanked by adaptor sequences were synthesized by Agilent Technologies, amplified by PCR and cloned into the pCRISPRia-v2 lentiviral vector (Addgene, 84832) using BstXI (NEB, R0113) and Blpl (NEB, R0585)97. .. The dCas9-ZIM3 plasmid was constructed using NEBuilder HiFi DNA Assembly Master Mix (NEB, E2621), the sequence for the ZIM3KRAB–dCas9 domain amplified from pLX303-ZIM3-KRAB-dCas9 (Addgene, 154472) by PCR with primers CM_oligo_1 and CM_oligo_2 (Supplementary Table 1) and the Lenti-SFFV-mCherry-dCas9-VP64 (ref. 46) (Addgene, 180263) lentiviral backbone digested with PmeI (NEB, R0560) and BamHI (NEB, R3136). .. For arrayed CRISPRi validation experiments, sgRNA sequences were ordered as Ultramers (IDT, 0.2 μM) with flanking adaptors, as in CM_oligo_3, cloned into the pCRISPRia-v2 lentiviral vector (Addgene, 84832) digested with Blpl (NEB, R0585) and BstXI (NEB, R0113) using NEBuilder HiFi DNA Assembly Master Mix (NEB, E2621) and transformed into STBL3 chemically competent cells (QB3 MacroLab).



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