Induced Pluripotent Stem Cell Derivation and Ex Vivo Gene Correction Using a Mucopolysaccharidosis Type 1 Disease Mouse Model
Joint Authors
Miki, Toshio
Garcia, Irving M.
Vazquez, Ludivina
Yanuaria, Lisa
Lopez, Omar
Ohashi, Kazuo
Rodriguez, Natalie S.
Source
Issue
Vol. 2019, Issue 2019 (31 Dec. 2019), pp.1-10, 10 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2019-04-01
Country of Publication
Egypt
No. of Pages
10
Abstract EN
Mucopolysaccharidosis type 1 (MPS-1), also known as Hurler’s disease, is a congenital metabolic disorder caused by a mutation in the alpha-L-iduronidase (IDUA) gene, which results in the loss of lysosomal enzyme function for the degradation of glycosaminoglycans.
Here, we demonstrate the proof of concept of ex vivo gene editing therapy using induced pluripotent stem cell (iPSC) and CRISPR/Cas9 technologies with MPS-1 model mouse cell.
Disease-affected iPSCs were generated from Idua knockout mouse embryonic fibroblasts, which carry a disrupting neomycin-resistant gene cassette (Neor) in exon VI of the Idua gene.
Double guide RNAs were used to remove the Neor sequence, and various lengths of donor templates were used to reconstruct the exon VI sequence.
A quantitative PCR-based screening method was used to identify Neor removal.
The sequence restoration without any indel mutation was further confirmed by Sanger sequencing.
After induced fibroblast differentiation, the gene-corrected iPSC-derived fibroblasts demonstrated Idua function equivalent to the wild-type iPSC-derived fibroblasts.
The Idua-deficient cells were competent to be reprogrammed to iPSCs, and pluripotency was maintained through CRISPR/CAS9-mediated gene correction.
These results support the concept of ex vivo gene editing therapy using iPSC and CRISPR/Cas9 technologies for MPS-1 patients.
American Psychological Association (APA)
Miki, Toshio& Vazquez, Ludivina& Yanuaria, Lisa& Lopez, Omar& Garcia, Irving M.& Ohashi, Kazuo…[et al.]. 2019. Induced Pluripotent Stem Cell Derivation and Ex Vivo Gene Correction Using a Mucopolysaccharidosis Type 1 Disease Mouse Model. Stem Cells International،Vol. 2019, no. 2019, pp.1-10.
https://search.emarefa.net/detail/BIM-1209413
Modern Language Association (MLA)
Miki, Toshio…[et al.]. Induced Pluripotent Stem Cell Derivation and Ex Vivo Gene Correction Using a Mucopolysaccharidosis Type 1 Disease Mouse Model. Stem Cells International No. 2019 (2019), pp.1-10.
https://search.emarefa.net/detail/BIM-1209413
American Medical Association (AMA)
Miki, Toshio& Vazquez, Ludivina& Yanuaria, Lisa& Lopez, Omar& Garcia, Irving M.& Ohashi, Kazuo…[et al.]. Induced Pluripotent Stem Cell Derivation and Ex Vivo Gene Correction Using a Mucopolysaccharidosis Type 1 Disease Mouse Model. Stem Cells International. 2019. Vol. 2019, no. 2019, pp.1-10.
https://search.emarefa.net/detail/BIM-1209413
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1209413