Integrated DNA Technologies (IDT) advances CRISPR research with the use of its gBlocks Gene Fragments. Detailed online in the latest issue of DECODED, CRISPRs can be used in conjunction with CRISPR associated (Cas) proteins, for example Cas9 has been well documented, to recognize and cleave complementary DNA.
This mechanism, which is found naturally within bacteria and archaea to protect cells from foreign sequence, has been harnessed for use in genome editing.
In most eukaryotes, the double strand break created is repaired by non-homologous end joining, which results in small INDELS that disrupt the genes; something which can be employed to achieve gene silencing at the genome level.
DNA repair mechanisms can incorporate foreign DNA into the cells’ genome through homologous recombination. The CRISPR/Cas9 system targets gene disruptions in a variety of organisms and cell types, and can insert sequence changes to targeted genomic regions.
Several recent papers have detailed the use of gBlocks Gene Fragments to create codon-optimized versions of Cas9 to facilitate efficient expression of inserted sequence in host cells.