LMNA co-regulated gene expression as a suitable readout after precise gene correction

Wang, H.; Krause, A.; Escobar, H.; Müthel, S.; Metzler, E.; Spuler, S.

LMNA co-regulated gene expression as a suitable readout after precise gene correction

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Item Type:	Article
Title:	LMNA co-regulated gene expression as a suitable readout after precise gene correction
Creators Name:	Wang, H., Krause, A., Escobar, H., Müthel, S., Metzler, E. and Spuler, S.
Abstract:	LMNA-related muscular dystrophy is an autosomal-dominant progressive disorder caused by mutations in LMNA. LMNA missense mutations are becoming correctable with CRISPR/Cas9-derived tools. Evaluating the functional recovery of LMNA after gene editing bears challenges as there is no reported direct loss of function of lamin A/C proteins in patient-derived cells. The proteins encoded by LMNA are lamins A/C, important ubiquitous nuclear envelope proteins but absent in pluripotent stem cells. We induced lamin A/C expression in induced pluripotent stem cells (iPSCs) of two patients with (LMNA)-related muscular dystrophy, NM_170707.4 (LMNA): c.1366A > G, p.(Asn456Asp) and c.1494G > T, p.(Trp498Cys), using a short three-day, serum-induced differentiation protocol and analyzed expression profiles of co-regulated genes, examples being COL1A2 and S100A6- We then performed precise gene editing of (LMNA) c.1366A > G using the near-PAMless (PAM: protospacer-adjacent motif) cytosine base editor. We show that the mutation can be repaired to 100% efficiency in individual iPSC clones. The fast differentiation protocol provided a functional readout and demonstrated increased lamin A/C expression as well as normalized expression of co-regulated genes. Collectively, our findings demonstrate the power of CRISPR/Cas9-mediated gene correction and effective outcome measures in a disease with, so far, little perspective on therapies.
Keywords:	Laminopathy, Muscular Dystrophy, LMNA Co-Regulated Genes, Near-PAMless Cytosine Base, Editor, Serum-Induced Differentiation (SID), Patient-Derived Induced Pluripotent Stem Cells (iPSCs)
Source:	International Journal of Molecular Sciences
ISSN:	1422-0067
Publisher:	MDPI
Volume:	23
Number:	24
Page Range:	15525
Date:	2 December 2022
Official Publication:	https://doi.org/10.3390/ijms232415525
PubMed:	View item in PubMed

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