A cost efficient protocol to introduce epitope tags by CRISPR-Cas9 mediated gene knock-in with asymmetric semi-double stranded template.

ABSTRACT: To study the biological function of uncharacterized proteins, specific antibodies are in high demand. However, the production of desirable antibodies such as highly specific or high affinity is not always successful. Furthermore, even if commercially available antibodies exist, the cost, quality, and accessibility often differ from country to country. In comparison, epitope tags are reliable and economical options since good antibodies against major epitope tags are commercially available. Although exogenously expressed epitope-tagged protein appears as a timely method, the excessive protein production may not faithfully recapitulate its biology. Thanks to the recent advances in genome editing by CRISPR-Cas9, HDR-mediated endogenous protein tagging has become an accessible approach for man

SUBMITTER: Fang Q 

PROVIDER: S-EPMC8374323 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature