Generation of endogenous BMP transcriptional reporter cells through CRISPR/Cas9 genome editing

Luke D. Hutchinson, Polyxeni Bozatzi, Thomas Macartney, Gopal P. Sapkota (Lead / Corresponding author)

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)peer-review

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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.

Original languageEnglish
Title of host publicationBone Morphogenetic Proteins
Subtitle of host publicationMethods and Protocols
EditorsMelissa B. Rogers
Place of PublicationNew York
PublisherHumana Press
Pages29-35
Number of pages7
Volume1891
ISBN (Electronic)9781493989041
ISBN (Print)9781493989034
DOIs
Publication statusPublished - 2018

Publication series

NameMethods in Molecular Biology
PublisherSpringer
Volume1891
ISSN (Print)1064-3745

Keywords

  • Transcription
  • Reporter vectors
  • CRISPR/Cas9
  • Genome editing

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