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URL: http://www.addgene.org/65779
Proper Citation: RRID:Addgene_65779
Insert Name: SaCas9 gRNA backbone, without spacer sequence
Organism: Synthetic
Bacterial Resistance: Ampicillin
Defining Citation: PMID:26098369
Vector Backbone Description: Backbone Marker:Joung lab (Addgene Plasmid # 43860); Vector Backbone:MLM3636; Vector Types:Mammalian Expression, CRISPR; Bacterial Resistance:Ampicillin
Comments: Use BsmBI sites to insert sgRNA spacer sequence into the vector. See the supplemental document "Joung Lab gRNA Cloning Protocol" for more details. UPDATE 5/31/2016: The Joung Lab recommends using BPK2660 (Addgene plasmid # 70709) instead of VVT1 as an SaCas9 guide RNA cloning vector, as guide RNAs cloned into BPK2660 are more effective than those cloned into VVT1. The difference in efficacy is due to VVT1 containing the full length guide RNA (predicted from the natural CRISPR sequence in S. aureus), whereas BPK2660 uses a truncated gRNA that is more effective. For the same oligo/spacer, the rate of mutagenesis is greater using BPK2660. See Kleinstiver et al., 2015 Nat Biotech for additional information on the truncated gRNA plasmid (https://www.ncbi.nlm.nih.gov/pubmed/26524662).
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Source: Addgene