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 over-expression or inhibition ofspecific genes hold a great deal ofpromise in terms of theirtherapeutic application. Here we optimized the direct PCR approach for rapid identification of genome-edited MSCs with only ten cells required, which reduces the time and labor to expand the MSC colonies. Combined with our previouslyoptimized guide RNA structure and plasmid construction strategy for Cas9, we successfully identified MSCcolonies over-expressing IL-10 in the AAVS1 locus.