High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells.
Fu Y, Foden JA, Khayter C, Maeder ML, Reyon D, Joung JK, Sander JD
Nat Biotechnol. 2013 Jun 23. doi: 10.1038/nbt.2623.
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PubMed
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Article
CRISPR-Cas/RGN expression plasmids for EGFP and Human genes
The paper describes the use of CRISPR-Cas/RNA-Guided Nuclease (RGN) technology to modify three different sites in the EGFP reporter gene (Addgene plasmid #47511–47513) and four different endogenous human genes (Addgene plasmid #47505–47510).
Note that gRNA produced from each of these vectors were used in conjunction with mRNA produced from the mammalian codon-optimized Cas9 expression vector (JDS246). Additional details can be found in the paper and users are encouraged to post questions to the Genome Engineering newsgroup (which can be joined here (Link opens in a new window)).
Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 47505 | pVC297 VEGF Site#1 | human gRNA expression vector targeting VEGF |
| 47506 | pVC299 VEGF Site#2 | human gRNA expression vector targeting VEGF |
| 47507 | pVC228 VEGF Site#3 | human gRNA expression vector targeting VEGF |
| 47508 | pFYF1548 EMX1 | human gRNA expression vector targeting EMX1 |
| 47509 | pDR366 RNF2 | human gRNA expression vector targeting RNF2 |
| 47510 | pDR348 FANCF | human gRNA expression vector targeting FANCF |
| 47511 | pFYF1320 EGFP Site#1 | human gRNA expression vector targeting EGFP |
| 47512 | pFYF1327 EGFP Site#2 | human gRNA expression vector targeting EGFP |
| 47513 | pFYF1328 EGFP Site#3 | human gRNA expression vector targeting EGFP |