Tools for Cancer Research: CRISPR/Cas-Based Gene Editing and Auxin-Induced Degron Systems

CRISPR/Cas9 and auxin-degron systems represent two powerful and complementary genetic tools that have revolutionized cancer research. The clustered regularly interspaced short palindromic repeats (CRISPR)-Cas (CRISPR-associated protein)-based gene editing system has been widely used in numerous biological fields, significantly enhancing the capacity of researchers to elucidate the underlying mechanisms of biological phenomena. The two most prevalent applications for CRISPR-mediated gene editing are knockout and knock-in. This protocol covers both single-gRNA (sgRNA) and dual-gRNA directed knockout systems. Furthermore, we describe the procedure for constructing an Auxin-Inducible Degron (AID) knock-in-mediated system to induce the rapid degradation of a target protein in cells, thereby investigating its function.