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- Complete kit for gene knockout via CRISPR (targeted sites around the 5’ end of the ORF)
- 2 guide RNA vectors in pCas-Guide to ensure an efficient cleavage
- Donor vector with predesigned homologous arms
- Knockin GFP-Puro for selection
- pCas-Guide-scramble is also provided as a negative control
- Knock-in GFP reporter for promoter study
- Knock-out genes at chromosomal level
Scheme of Genome-editing knockout kit using CRISPR
The latest tool in genome editing – CRISPR/Cas9 – allows for specific genome disruption and replacement in a flexible and simple system resulting in high specificity and low cell toxicity. The CRISPR/Cas9 genome editing system requires the co- expression of a Cas9 protein with a guide RNA vector expressed from the human U6 polymerase III promoter. With the protospacer-adjacent motif (PAM - the sequence NGG) present at the 3′ end, Cas9 will unwind the DNA duplex and cleave both strands upon recognition of a target sequence by the guide RNA.
- Multiplex Genome Engineering Using CRISPR/Cas Systems. Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, Hsu PD, Wu X, Jiang W, Marraffini LA, Zhang F.
Science 2013 Feb 15;339(6121):819-23
- RNA-Guided Human Genome Engineering via Cas9. Mali P, Yang L, Esvelt KM, Aach J, Guell M, DiCarlo JE, Norville JE, Church GM
Science 2013 Feb 15;339(6121):823-6