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Disease relevant mutations can be recapitulated or even correct by introducing a point mutation with a single base change, or by inserting/deleting large gene sequences. These endogenous knock-in cell lines enable a clear understanding of the contribution of the gene or mutation to a phenotype. There are unlimited uses for point mutation cell lines at pre-defined loci, including functional assays, drug screening, FACS screening of membrane proteins, gene expression studies, gene therapy and antibody immunization boosting research.
The RNA-guided DNA endonuclease Cas9 associates with a synthetic single guide RNA (gRNA) and cleaves double-stranded DNA targets complementary to the guide RNA. The double-stranded break occurs 3 bp upstream of the PAM site, allowing targeted sequence modifications via homology-directed repair (HDR) pathway for precise insertion of point mutations at the targeted sites. If the repair of that double strand break occurs via the homology-directed repair pathway, the donor sequence can be inserted and a single base change is introduced in to that gene.
Creative Biogene’s experienced scientists will provide researchers with a reliable, affordable, and fast knock-in cell line generation service, which includes gRNA/donor DNA construction, PCR sequencing, and engineered cell line generation. Our mammalian cell lines with long-term gene editing can be one of the most effective tools in your research or drug discovery program.
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