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 (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 |