dc.contributor.author
Wang, Haicui
dc.contributor.author
Krause, Anne
dc.contributor.author
Escobar, Helena
dc.contributor.author
Müthel, Stefanie
dc.contributor.author
Metzler, Eric
dc.contributor.author
Spuler, Simone
dc.date.accessioned
2023-09-22T13:10:04Z
dc.date.available
2023-09-22T13:10:04Z
dc.identifier.uri
https://refubium.fu-berlin.de/handle/fub188/40950
dc.identifier.uri
http://dx.doi.org/10.17169/refubium-40671
dc.description.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.
en
dc.rights.uri
https://creativecommons.org/licenses/by/4.0/
dc.subject
muscular dystrophy
en
dc.subject
LMNA co-regulated genes
en
dc.subject
near-PAMless cytosine base editor
en
dc.subject
serum-induced differentiation (SID)
en
dc.subject
patient-derived induced pluripotent stem cells (iPSCs)
en
dc.subject.ddc
600 Technik, Medizin, angewandte Wissenschaften::610 Medizin und Gesundheit::610 Medizin und Gesundheit
dc.title
LMNA Co-Regulated Gene Expression as a Suitable Readout after Precise Gene Correction
dc.type
Wissenschaftlicher Artikel
dcterms.bibliographicCitation.articlenumber
15525
dcterms.bibliographicCitation.doi
10.3390/ijms232415525
dcterms.bibliographicCitation.journaltitle
International Journal of Molecular Sciences
dcterms.bibliographicCitation.number
24
dcterms.bibliographicCitation.originalpublishername
MDPI
dcterms.bibliographicCitation.volume
23
refubium.affiliation
Charité - Universitätsmedizin Berlin
refubium.resourceType.isindependentpub
no
dcterms.accessRights.openaire
open access
dcterms.bibliographicCitation.pmid
36555163
dcterms.isPartOf.eissn
1422-0067