pCC_10 - hU6-BsmBI-sgRNA(E+F)-barcode-EFS-KRAB-dCas9NG-NLS-2A-Puro-WPRE
(Plasmid
#139095)
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PurposeExpresses human codon-optimized inactive SpCas9-NG fused to a transcriptional repressor KRAB in mammalian cells. For cloning of sgRNAs using BsmBI. Contains a barcode downstream of sgRNA cassette.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 139095 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonelentiCRISPRv2
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Vector typeMammalian Expression, Lentiviral, CRISPR
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Selectable markersPuromycin
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)NEB Stable
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameKRAB-dSpCas9-NG
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SpeciesStaphylococcus pyogenes
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MutationD10A, H840A, L1111R, D1135V,G1218R, E1219F, A1322R, R1335V, T1337R
Cloning Information
- Cloning method Gibson Cloning
Resource Information
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Supplemental Documents
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
pCC_10 - hU6-BsmBI-sgRNA(E+F)-barcode-EFS-KRAB-dCas9NG-NLS-2A-Puro-WPRE was a gift from Neville Sanjana (Addgene plasmid # 139095 ; http://n2t.net/addgene:139095 ; RRID:Addgene_139095) -
For your References section:
High-Throughput Screens of PAM-Flexible Cas9 Variants for Gene Knockout and Transcriptional Modulation. Legut M, Daniloski Z, Xue X, McKenzie D, Guo X, Wessels HH, Sanjana NE. Cell Rep. 2020 Mar 3;30(9):2859-2868.e5. doi: 10.1016/j.celrep.2020.02.010. 10.1016/j.celrep.2020.02.010 PubMed 32130891