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Purpose(Empty Backbone) Plasmid with Cas9, two U6 promoters and two gRNA scaffolds that allows for inserting two gRNAs for combinatorial CRISPR Screen or double-knock-out (DKO) screen
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Depositing Lab
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Sequence Information
Ordering
This material is available to academics and nonprofits only.
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 183192 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 | |
Backbone
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Vector backbonelentiCRISPR v2
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Backbone manufacturerPlasmid #52961
- Backbone size (bp) 14873
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Modifications to backboneAdded another U6 promotor and scaffold with three enzyme cutting sites
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Vector typeMammalian Expression, Lentiviral, CRISPR
- Promoter hU6, mU6
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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
Cloning Information
- Cloning method Gibson Cloning
- 5′ sequencing primer hU6-F (Common Sequencing Primers)
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.
How to cite this plasmid
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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:
lentiCRISPR_DKO was a gift from Lang Li (Addgene plasmid # 183192 ; http://n2t.net/addgene:183192 ; RRID:Addgene_183192) -
For your References section:
Generation of dual-gRNA library for combinatorial CRISPR screening of synthetic lethal gene pairs. Tang S, Wu X, Liu J, Zhang Q, Wang X, Shao S, Gokbag B, Fan K, Liu X, Li F, Cheng L, Li L. STAR Protoc. 2022 Aug 24;3(3):101556. doi: 10.1016/j.xpro.2022.101556. eCollection 2022 Sep 16. 10.1016/j.xpro.2022.101556 PubMed 36060092
