CRISPR CAS9 Genome Editing

Overview

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. The functional cassette synthesized by Blue Heron in the rescue donor vector can then be inserted into the unwound DNA. The repaired genome will now express your desired sequence with or without tags.

Cas9 Further Reading

CRISPR/CAS9 Custom Services

 

Custom Services Reference Price
Target sequence cloning into CRISPR/Cas9 vectors $345
$150
Donor vector construction with a predesigned cassette ~$880
1.2kb of homologous arms (left + right)
Donor vector of your own design Gene synthesis cost, $0.4-0.5/bp

diagram-gene-editing-recombiiPredesigned Cassette

  • A ptGFPPuro
  • B pRFPBSD
  • C pLucPuro
  • D ptGFPPuroRev
  • E BFPneo