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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. and Krause, A. and Escobar, H. and Müthel, S. and 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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