Factor-induced Reprogramming and Zinc Finger Nuclease-aided Gene Targeting Cause Different Genome Instability in β-Thalassemia Induced Pluripotent Stem Cells (iPSCs).

The Journal of Biological Chemistry
Ning MaGuangjin Pan

Abstract

The generation of personalized induced pluripotent stem cells (iPSCs) followed by targeted genome editing provides an opportunity for developing customized effective cellular therapies for genetic disorders. However, it is critical to ascertain whether edited iPSCs harbor unfavorable genomic variations before their clinical application. To examine the mutation status of the edited iPSC genome and trace the origin of possible mutations at different steps, we have generated virus-free iPSCs from amniotic cells carrying homozygous point mutations in β-hemoglobin gene (HBB) that cause severe β-thalassemia (β-Thal), corrected the mutations in both HBB alleles by zinc finger nuclease-aided gene targeting, and obtained the final HBB gene-corrected iPSCs by excising the exogenous drug resistance gene with Cre recombinase. Through comparative genomic hybridization and whole-exome sequencing, we uncovered seven copy number variations, five small insertions/deletions, and 64 single nucleotide variations (SNVs) in β-Thal iPSCs before the gene targeting step and found a single small copy number variation, 19 insertions/deletions, and 340 single nucleotide variations in the final gene-corrected β-Thal iPSCs. Our data revealed that substantia...Continue Reading

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Citations

Jan 15, 2016·Molecular Therapy : the Journal of the American Society of Gene Therapy·Versha PrakashRafael J Yáñez-Muñoz
Feb 16, 2018·Human Gene Therapy·Yang YangYuquan Wei
Feb 28, 2018·Stem Cell Research & Therapy·Methichit WattanapanitchSurapol Issaragrisil
Sep 10, 2019·Annals of Hematology·Xie YingjunSun Xiaofang
Sep 9, 2020·BioDrugs : Clinical Immunotherapeutics, Biopharmaceuticals and Gene Therapy·Karine Sii-FeliceEmmanuel Payen
Jun 7, 2019·Disease Models & Mechanisms·Maria Georgomanoli, Eirini P Papapetrou
Jan 5, 2017·The Korean Journal of Internal Medicine·Eun Ji KimJi Hyeon Ju
Sep 12, 2020·Stem Cell Research & Therapy·Hala GabrMaha Hamdi El Sissy
Sep 25, 2017·Nature Communications·Yongli ShanGuangjin Pan

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