Chen and Wente Lab CRISPR Plasmids Available from Addgene
We have synthesized a Cas9 coding sequence with nuclear localization signals (nls) at both its amino and carboxyl termini and codons optimized for zebrafish expression (nls-zCas9-nls). Injection of in vitro transcribed nls-zCas9-nls mRNA with a gene-specific chimeric guide RNA consistently resulted in high mutagenic efficiency in both somatic and germline cells. Injected fish often exhibit loss-of-function phenotype, indicating prevalent biallelic inactivation. The injected fish produces predominantly mutation-carrying progeny. In addition, multiple loci can be efficiently targeted simultaneously in the same fish.
A protocol for synthesizing gRNAs: gRNA plasmid construction protocol (66.8 KB)
These plasmids are descirbed in:
Efficient multiplex biallelic zebrafish genome editing using a CRISPR nuclease system. Jao LE, Wente SR, Chen W. Proc Natl Acad Sci . 2013 Aug 25. PubMed .
Individual plasmids can be ordered via the links below:
|46761||pT7tyrgRNA : A gRNA targeting Tyr.||Add to Cart|
|46760||pT7EGFPgRNA : A gRNA targeting EGFP.||Add to Cart|
|46757||pT3TS-nCas9n : A Cas9 expression plasmid.||Add to Cart|
|46759||pT7-gRNA : A gRNA core.||Add to Cart|
|47929||pCS2-nCas9n : For expression of an optimized Cas9 for genome-editing in zebrafish.||Add to Cart|
|47930||pT7goldRNA : A plasmid for in vitro transcription of golden gRNA.||Add to Cart|
|47931||pT7mitfagRNA :A plasmid for in vitro transcription of mitfa gRNA.||Add to Cart|
|47932||pT7ddx19gRNA : A plasmid for in vitro transcription of ddx19 gRNA.||Add to Cart|