CRISPR/Cas9 Genome Editing
RNA-guided genome editing by CRISPR/Cas9 allows for specific genome
disruption and replacement in a flexible and simple system. This system
requires the co- expression of a Cas9 protein with a guide RNA vector.
The Cas9 protein is an RNA-guided endonuclease that catalyzes
site-specific cleavage of double stranded DNA. The cleavage site is
within the target sequence 3 bases upstream of the protospacer-adjacent-motif
(PAM - the sequence NGG). Then, the double-strand break is repaired by
homologous recombination with the modified genome template from the
rescue donor vector. In this way, we can generate insertions, deletion,
point mutant, in-frame GFP fusions, or any other modification.
|Target sequence design and cloning into
|Donor vector construction with a
|Donor vector of your own design gene
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