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The RNA-guided CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR associated nuclease 9) system is a revolutionary genome editing tool derived from the bacterial Type II CRISPR adaptive immune system. Its primary function in bacteria is the endonucleolytic destruction of invading plasmid or phage DNA. In eukaryotic systems, CRISPR/Cas9 is used for genomic editing through specific targeting of DNA by a single guide RNA (sgRNA), a combination of the CRISPR RNA (crRNA) and the trans-activating crRNA (tracrRNA), mediated through base pairing over the ~20-nt guide sequence. Cas9 recognizes a very short conserved sequence (a few nucleotides in length) adjacent to the guide sequence called the “protospacer adjacent motif” (PAM).
Once directed to the DNA target site, Cas9 generates a double-strand break (DSB) that can be repaired either through the indel mutation-introducing non-homologous end-joining (NHEJ) or the high-fidelity homologous directed repair (HDR), resulting in gene knockout effects or template-dependent gene replacement. GeneTex engages CRISPR/Cas9 technology to validate antibody specificity At GeneTex, our immediate goal is to integrate CRISPR/Cas9 technology into our quality assurance process as the “knockout” arm of our antibody specificity validation efforts. In a collaborative effort with EdiGene, a company that generates knockout or genome-edited cell lines using CRISPR/Cas9- and TALEN-mediated methodologies, GeneTex will use lysates made from these cell lines for western blot assays to confirm target specificity of our antibodies. Our hope is to continue providing the research community with the highest quality and most stringently validated antibodies supported by these definitive KO data.