Amsbio has introduced the CRISPR/Cas9 - a full gene editing tool designed to produce a genetically modified cell using any mammalian cell line and targeting any gene.
The tool works within the field of genomic editing by providing scientists with a powerful tool able to change any gene, in any cell in a highly targeted manner and without introducing foreign DNA.
The benefits of CRISPR/Cas9 over previous forms of gene editing, such as TALENs and zinc finger, are that it is much simpler to implement and has higher efficiency at performing bi-allelic gene modifications.
Using CRISPPR/Cas9 technology it is now possible to create knock-in, knock-outs and mutations of any gene in any cell line.
CRISPR/Cas9 allows for specific genome disruption and replacement in a flexible and simple system resulting in high specificity and low cell toxicity.
The 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 synthesised 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.
