Generation of Endogenous BMP Transcriptional Reporter Cells Through CRISPR/Cas9 Genome Editing.

Generation of Endogenous BMP Transcriptional Reporter Cells Through CRISPR/Cas9 Genome Editing.
Methods Mol Biol. 2019;1891:29-35
Authors: Hutchinson LD, Bozatzi P, Macartney T, Sapkota GP
Abstract
Transcriptional reporter systems allow researchers to investigate the function and regulation of transcription factors. Conventional systems employ artificial cDNA overexpression vectors containing either a promoter fragment or specific nucleotide sequence repeats upstream of firefly luciferase or fluorescent reporters, such as green fluorescence protein (GFP) cDNA. These systems suffer mainly from the lack of chromatin context. Here, we describe the rapid generation of endogenous transcriptional reporter cells for the bone morphogenetic protein (BMP) pathway using CRISPR/Cas9 genome editing. In principle, our methodology can be applied to any cell line. The endogenous reporters will provide a robust system for the investigation of BMP transcriptional activity in the context of native chromatin landscape and facilitate chemical and genetic screens.
PMID: 30414124 [PubMed - in process]

Source: Mol Biol Cell - November 12, 2018 Category: Molecular Biology Authors: Hutchinson LD, Bozatzi P, Macartney T, Sapkota GP Tags: Methods Mol Biol Source Type: research