CRISPR 101: Making a Knock-In Cell Line
Type
Blog Post
...Disable the other repair mechanisms
Mammalian cells have 2 primary repair mechanisms besides HR - Non-homologous... However, if you have a larger knock-in (200 bp – 2 kb), a dsDNA donor will need to be used due to synthesis...accidentally change your protein of interest.
Fig. 2: Stop re-cutting with PAM and guide RNA disrupting.... Com., 8(2024), (2017). 10.1038/s41467-017-01836-2
Charpentier, M., Khedher, A. H. Y., Menerot, S., Brion...varying across cell types. Variance can be caused by cell cycle rates; rapidly diving cells will have higher... of your cells.
Enrich S-phase cells
HR operates predominantly in S and G2 phase of the cell cycle, when...HDR events, ensure cells are actively cycling through the cell cycle stages. If cells are overly confluent...