Generation of gene-corrected iPSC line, KIOMi002-A, from Parkinson's disease patient iPSC with LRRK2 G2019S mutation using BAC-based homologous recombination

doi:10.1016/j.scr.2019.101649

Stem Cell Research

Volume 41, December 2019, 101649

https://doi.org/10.1016/j.scr.2019.101649 Get rights and content

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Highlights

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We genetically corrected the LRRK2 G2019S gene mutation.
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A BAC-based homologous recombination system was used as a gene replacement tool.
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Gene-corrected iPSCs had the characteristics of pluripotent stem cells.

Abstract

Mutations in leucine-rich repeat kinase 2 (LRRK2) gene (LRRK2 G2019S) is a representative autosomal dominant mutation that can cause Parkinson's disease (PD). A bacterial artificial chromosome-based homologous recombination (BAC-based HR) system was utilized for gene therapy of LRRK2 G2019S-mutant induced pluripotent stem cells (iPSCs) produced by reprogramming episomal vectors. The gene-corrected iPSCs retained typical pluripotency required for their spontaneous differentiation into differentiated cells. The iPSCs had a normal karyotype and were confirmed to have no off-target sites by melting curve analysis.