URL: http://www.addgene.org/104993
Proper Citation: RRID:Addgene_104993
Bacterial Resistance: Ampicillin
Defining Citation: PMID:30894629
Vector Backbone Description: Backbone Marker:Brett Stringer from Feng Zhang plasmid lentiCRISPR v2 (Addgene plasmid #52961); Backbone Size:10437; Vector Backbone:lentiCRISPRv2 blast (Addgene #98293); Vector Types:Mammalian Expression, Lentiviral, CRISPR; Bacterial Resistance:Ampicillin
Comments: This 3rd generation lentiviral plasmid expresses a S. pyogenes CRISPR chimeric RNA element with a customizable sgRNA from a U6 promoter and blasticidin resistance gene from an EF-1a core promoter. This plasmid does NOT contain Cas9. It should be used in conjunction with lentiCas9 puro, lentiCas9 hygro or lentiCas9 neo or with cell lines expressing Cas9. After cloning a double-stranded oligonucleotide specifying a gRNA between the two BsmBI restriction sites, use hU6-F (5'-GAGGGCCTATTTCCCATGATT-3') or LKO.1 5’(5'- GACTATCATATGCTTACCGT-3') to sequence the chimeric RNA element. Instruction for gRNA double-stranded oligonucleotide design and cloning can be found in the Zhang lab protocol, https://media.addgene.org/data/plasmids/52/52961/52961-attachment_B3xTwla0bkYD.pdf
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Source: Addgene