Point Mutation Stable Cell Line Construction Services

Seattle Genova’s experienced scientists will give researchers a reliable, affordable, and fast knock-in cell line generation service, which contains 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 largely beneficial tools in your research or drug discovery program.

Point mutations are the type of mutations in which a nitrogen base in a DNA strand is either added or deleted or displaced with another resulting in a difference in codon hence a change in the protein. To know what a point mutation is, it must first be understood that DNA molecules—and hence the genes found along their length—are organized of building blocks called nucleotide bases. A proto-oncogene may be altered into an oncogene through a single alteration of a nucleotide. 

Disease-relevant mutations can be recapitulated or even corrected by initiating a point mutation with a single base difference, or by inserting/deleting large gene sequences. These endogenous knock-in cell lines facilitate a clear awareness of the assistance of the gene or mutation to a phenotype. There are infinite usages for point mutation cell lines at pre-defined loci, containing functional assays, drug screening, FACS screening of membrane proteins, gene expression studies, gene therapy and antibody immunization boosting studies.

The RNA-guided DNA endonuclease Cas9 assists with a synthetic single guide RNA (gRNA) and cleaves double-stranded DNA targets complementary to the direction RNA. The double-stranded break occurs 3 bp upstream of the PAM site, allowing targeted sequence modifications via homology-directed repair (HDR) pathway for critical insertion of point mutations at the targeted areas. If the repair of that double-strand break exists via the homology-directed repair pathway, the donor sequence can be inserted and a single base alteration is introduced into that gene.