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Overview Cas9-cell line gene mutation (CGM) stands as a highly effective tool for investigating cell gene functions, particularly enabling the in-situ study of point mutations within cells.
Key Advancements
Our achievements With a track record of over 100 successful cases, Hysigen has demonstrated proficiency in transfecting commonly used cell lines, including Jurkat, NK-92, BV2, C2C12, EMT6, B16-F10, ARPE-19, HepG2, THP-1, HCT116, A549, RAW264.7, MDA-MB-231, MDA-MB-468, 4T1, and more. Workflow * We kindly remind you that we provide gene editing services for primary cells, stem cells or iPS cells.
CRISPR-mediated point mutation CRISPR-mediated point mutation involves using the CRISPR-Cas9 system to introduce precise changes in a single nucleotide of a targeted gene. The process begins with the design of a guide RNA (gRNA) that directs the Cas9 enzyme to the specific genomic location where the point mutation is desired. The Cas9 protein acts as molecular scissors, creating a double-strand break (DSB) at the target site. The cell's natural repair mechanism, Homology Directed Repair (HDR), then comes into play. HDR can be employed with a donor DNA template containing the desired point mutation. This template guides the precise repair of the DNA, incorporating the desired mutation at the targeted nucleotide position. The CRISPR-mediated point mutation mechanism enables researchers to engineer specific changes in the genome with remarkable precision. Point mutation of JAK2 in MEG-01 Cells Using CRISPR/Cas9
Experimental design Strategy Identification results PCR screening Final Clone Sequences #4C7: AAGCAGCAAGTATGATGAGCAAGCTTTCTCACAAGCATTTGGTTTTAAATTATGGAGTATGTTTCTGTGGAGACGAGAGTAAGTAAAACTACAGGCTTTCTAATGCCTTTCTCAGAGCAT * Point mutation: V617F |