Control of Adipogenic Differentiation in Mesenchymal Stem Cells via Endogenous Gene Activation Using CRISPR-Cas9

Mesenchymal stem
cells (MSCs) are of interest in regenerative medicine
owing to their multilineage differentiation and self-renewal properties.
Understanding the in vivo differentiation process
is necessary for clinical applications including cell therapy and
transplantation. This remains challenging owing to the lack of induction
methods that imitate the natural programming process. Endogenous gene
regulation of tissue-specific transcription factors is therefore desirable.
In the present study, we demonstrated endogenous activation of adipogenic
genes through the dCas9-based transcription system and achieved efficient
induction of different types of adipocyte-like cells from MSCs. Interestingly,
the MSCs converted via single-gene activation exhibited morphological
and molecular properties of white adipocytes, while beige adipocyte-like
cells were induced via multiplex gene activation of three specific
transcription factors. These results reveal that the fate of MSCs
can be effectively manipulated by direct activation of specific endogenous
gene expression using a dCas9-based activator with reduced exogenous
additives.