Rapid generation of endogenously driven transcriptional reporters in cells through CRISPR/Cas9.

ABSTRACT: CRISPR/Cas9 technologies have been employed for genome editing to achieve gene knockouts and knock-ins in somatic cells. Similarly, certain endogenous genes have been tagged with fluorescent proteins. Often, the detection of tagged proteins requires high expression and sophisticated tools such as confocal microscopy and flow cytometry. Therefore, a simple, sensitive and robust transcriptional reporter system driven by endogenous promoter for studies into transcriptional regulation is desirable. We report a CRISPR/Cas9-based methodology for rapidly integrating a firefly luciferase gene in somatic cells under the control of endogenous promoter, using the TGFβ-responsive gene PAI-1. Our strategy employed a polycistronic cassette containing a non-fused GFP protein to ensure the detection of tr

SUBMITTER: Rojas-Fernandez A 

PROVIDER: S-EPMC4413877 | biostudies-literature | 2015 Apr

REPOSITORIES: biostudies-literature