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A Cas9 Variant for Efficient Generation of Indel-Free Knockin or Gene-Corrected Human Pluripotent Stem Cells.

Howden SE, McColl B, Glaser A, Vadolas J, Petrou S, Little MH, Elefanty AG, Stanley EG
Stem Cell Reports. 2016 Sep 13;7(3):508-17. doi: 10.1016/j.stemcr.2016.07.001. Epub 2016 Aug 4. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
80424pDNR-SpCas9-GemUsed for in vitro transcription of SpCas9-Gem mRNA. Cas9 is fused to Geminin peptide and is degraded during G0/G1 phases of the cell-cycle to minimize indels caused by non-homologous end joining
80425pDNR-SpCas9Used for in vitro transcription of SpCas9 mRNA.
80426pDNR-SpCas9-Cdt1Used for in vitro transcription of SpCas9-Cdt1 mRNA. Cas9 is fused to Cdt1 peptide and is degraded during S/G2 phases of the cell-cycle.
80427pSMART-sgRNA (Sp)U6 promoter driven sgRNA expression plasmid. Annealed oligonucleotides encoding crRNA sequence are ligated into BbsI site.

Antibodies from Article