Journal article

Generating an iPSC line (with isogenic control) from the PBMCs of an ACTA1 (p.Gly148Asp) nemaline myopathy patient

Peter J Houweling, Chantal A Coles, Chrystal F Tiong, Bridget Nielsen, Alison Graham, Penny McDonald, Annabelle Suter, Adam T Piers, Robin Forbes, Monique M Ryan, Sara E Howden, Shireen R Lamande, Kathryn N North

STEM CELL RESEARCH | ELSEVIER | Published : 2021

Abstract

To produce an in vitro model of nemaline myopathy, we reprogrammed the peripheral blood mononuclear cells (PBMCs) of a patient with a heterozygous p.Gly148Asp mutation in exon 3 of the ACTA1 gene to iPSCs. Using CRISPR/Cas9 gene editing we corrected the mutation to generate an isogenic control line. Both the mutant and control show a normal karyotype, express pluripotency markers and could differentiae into the three cell states that represent embryonic germ layers (endoderm, mesoderm and neuroectoderm) and the dermomyotome (precursor of skeletal muscle). When differentiated these cell lines will be used to explore disease mechanisms and evaluate novel therapeutics.

Grants

Funding Acknowledgements

The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Peter Houweling reports financial support was provided by Murdoch Childrens Research Institute. Peter Houweling reports a relationship with Murdoch Childrens Research Institute that includes: employment.