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PurposedCas9 with a C-term NLS-KRAB(Kox1)-MeCP2 fusion (no linker), and GFP linked by a P2A site from a SFFV promoter with an upstream ubiquitous chromatin opening element
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Depositing Labs
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 188902 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepMH0006, Addgene # 135448
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Vector typeLentiviral
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Selectable markersEGFP
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 namedCas9-KRAB(Kox1)-MeCP2
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SpeciesMultiple
- Promoter SFFV
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Tags
/ Fusion Proteins
- NLS-KRAB(Kox1)-MeCP2 fusion (no linker) (C terminal on insert)
- P2A-GFP (C terminal on insert)
Cloning Information
- Cloning method Gibson Cloning
Resource Information
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Supplemental Documents
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Article Citing this Plasmid
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.
Depositor Comments
Human lentiviral vector for expression of dCas9 with a C-terminal HA-2xNLS-KRAB(Kox1)-MeCP2 fusion (no linker), and GFP linked by a P2A site from a SFFV promoter with an upstream ubiquitous chromatin opening element. pCL51. Please visit https://doi.org/10.1101/2022.07.13.499814 for bioRxiv preprint.
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:
pHR-UCOE-SFFV-dCas9-KRAB(Kox1)-MeCP2-P2A-EGFP was a gift from Marco Jost & Jonathan Weissman (Addgene plasmid # 188902 ; http://n2t.net/addgene:188902 ; RRID:Addgene_188902) -
For your References section:
Maximizing CRISPRi efficacy and accessibility with dual-sgRNA libraries and optimal effectors. Replogle JM, Bonnar JL, Pogson AN, Liem CR, Maier NK, Ding Y, Russell BJ, Wang X, Leng K, Guna A, Norman TM, Pak RA, Ramos DM, Ward ME, Gilbert LA, Kampmann M, Weissman JS, Jost M. eLife. 2022 Dec 28;11:e81856. doi: 10.7554/eLife.81856. 10.7554/eLife.81856 PubMed 36576240