Rapid identification of genome-edited mesenchymal stem cell colonies via Cas9

Abstract

Mesenchymal stem cells (MSCs) have been intensively investigated and widely applied in regenerative medicine and immune modulation. However, their efficacy declines during the aging or disease process. Thus, genome-edited MSCs with overexpression or inhibition of specific genes hold a great deal of promise in terms of their therapeutic application. Here we optimized the direct PCR approach for rapid identification of genome-edited MSCs with only ten cells, which reduces the time and labor required to expand the MSC colonies. Combining with our previously optimized guide RNA structure and plasmid construction strategy for Cas9, we successfully identified MSC colonies overexpressing IL-10 in the AAVS1 locus.