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 (RRID:SCR_002037)
Description: Non-profit plasmid repository dedicated to helping scientists around the world share high-quality plasmids. Facilitates archiving and distributing DNA-based research reagents and associated data to scientists worldwide. Repository contains over 65,000 plasmids, including special collections on CRISPR, fluorescent proteins, and ready-to-use viral preparations. There is no cost for scientists to deposit plasmids, which saves time and money associated with shipping plasmids themselves. All plasmids are fully sequenced for validation and sequencing data is openly available. We handle the appropriate Material Transfer Agreements (MTA) with institutions, facilitating open exchange and offering intellectual property and liability protection for depositing scientists. Furthermore, we curate free educational resources for the scientific community including a blog, eBooks, video protocols, and detailed molecular biology resources.
URL: http://www.addgene.org