Generation of a human ACTA1-tdTomato reporter iPSC line using CRISPR/Cas9 editing

Peter J Houweling; Vanessa Crossman; Chrystal F Tiong; Chantal A Coles; Rhonda L Taylor; Joshua S Clayton; Alison Graham; Katerina Vlahos; Sara E Howden; Kathryn N North

doi:10.1016/j.scr.2024.103313

Generation of a human ACTA1-tdTomato reporter iPSC line using CRISPR/Cas9 editing

Stem Cell Res. 2024 Mar:75:103313. doi: 10.1016/j.scr.2024.103313. Epub 2024 Jan 23.

Affiliations

¹ Murdoch Children's Research Institute, Melbourne, Victoria, Australia; Department of Pediatrics, The University of Melbourne, Victoria, Australia. Electronic address: Peter.houweling@mcri.edu.au.
² Murdoch Children's Research Institute, Melbourne, Victoria, Australia.
³ Murdoch Children's Research Institute, Melbourne, Victoria, Australia; Department of Pediatrics, The University of Melbourne, Victoria, Australia.
⁴ Harry Perkins Institute of Medical Research, QEII Medical Centre, Nedlands, WA, Australia; Centre for Medical Research, University of Western Australia, QEII Medical Centre, Nedlands, WA, Australia.

PMID: 38277710
DOI: 10.1016/j.scr.2024.103313

Abstract

We used gene editing to introduce DNA sequences encoding the tdTomato fluorescent protein into the α -skeletal actin 1 (ACTA1) locus to develop an ACTA1-tdTomato induced pluripotent stem cell reporter line for monitoring differentiation of skeletal muscle. This cell line will be used to better understand skeletal muscle maturation and development in vitro as well as provide a useful tool for drug screening and the evaluation of novel therapeutics for the treatment of skeletal muscle disease.

MeSH terms

Actins / genetics
Actins / metabolism
CRISPR-Cas Systems* / genetics
Humans
Induced Pluripotent Stem Cells* / metabolism
Muscle, Skeletal / metabolism
Red Fluorescent Protein*

Substances

Actins
tdTomato
Red Fluorescent Protein