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URL: http://www.addgene.org/199223
Proper Citation: RRID:Addgene_199223
Insert Name: Expression unit for mCherry - monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)
Organism: Synthetic
Bacterial Resistance: Ampicillin
Defining Citation: PMID:36928106
Vector Backbone Description: Backbone Size:2700; Vector Backbone:pBR322; Vector Types:Mammalian Expression, Gene targeting donor plasmid; Bacterial Resistance:Ampicillin
Comments: This target site-containing donor plasmid can be used to insert a transgene at the human CCR5 safe harbor locus when delivered together with gRNA expression plasmid AY22_pgRNA.R5.1 (Addgene plasmid #100294) and a SpCas9 nuclease (HMEJ-based gene targeting) or a SpCas9.D10A nickase (ITPN-based gene targeting). Note: to reduce the chances for off-targeting donor DNA insertions when applying HMEJ-based genome editing it is preferable using AW01_pU.CAG.eSpCas9(1.1).rBGpA (Addgene plasmid #199255) or a similar construct, i.e., expressing a high specificity nuclease. Combining this donor with a S. pyogenes Cas9 nickase (e.g., Addgene plasmids #199256, #199253, #199254 or #199252) leads to seamless ITPN-mediated gene targeting.
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Source: Addgene