Bidirectional manipulation of gene expression in adipocytes using CRISPRa and siRNA.

ABSTRACT: OBJECTIVE:Functional investigation of novel gene/protein targets associated with adipocyte differentiation or function heavily relies on efficient and accessible tools to manipulate gene expression in adipocytes in vitro. Recent advances in gene-editing technologies such as CRISPR-Cas9 have not only eased gene editing but also greatly facilitated modulation of gene expression without altering the genome. Here, we aimed to develop and validate a competent in vitro adipocyte model of controllable functionality as well as multiplexed gene manipulation in adipocytes, using the CRISPRa "SAM" system and siRNAs to simultaneously overexpress and silence selected genes in the same cell populations. METHODS:We introduced a stable expression of dCas9-VP64 and MS2-P65, the core components of the CRIPS

SUBMITTER: Lundh M 

PROVIDER: S-EPMC5641601 | biostudies-literature | 2017 Oct

REPOSITORIES: biostudies-literature