Functional Correction of Large Factor VIII Gene Chromosomal Inversions in Hemophilia A Patient-Derived iPSCs Using CRISPR-Cas9

Chul Yong Park, Duk Hyoung Kim, Jeong Sang Son, Jin Jea Sung, Jaehun Lee, Sangsu Bae, Jong-Hoon Kim, Dong Wook Kim, Jin Soo Kim

Research output: Contribution to journalArticle

132 Citations (Scopus)

Abstract

Summary Hemophilia A is an X-linked genetic disorder caused by mutations in the F8 gene, which encodes the blood coagulation factor VIII. Almost half of all severe hemophilia A cases result from two gross (140-kbp or 600-kbp) chromosomal inversions that involve introns 1 and 22 of the F8 gene, respectively. We derived induced pluripotent stem cells (iPSCs) from patients with these inversion genotypes and used CRISPR-Cas9 nucleases to revert these chromosomal segments back to the WT situation. We isolated inversion-corrected iPSCs with frequencies of up to 6.7% without detectable off-target mutations based on whole-genome sequencing or targeted deep sequencing. Endothelial cells differentiated from corrected iPSCs expressed the F8 gene and functionally rescued factor VIII deficiency in an otherwise lethal mouse model of hemophilia. Our results therefore provide a proof of principle for functional correction of large chromosomal rearrangements in patient-derived iPSCs and suggest potential therapeutic applications.

Original languageEnglish
Article number1805
Pages (from-to)213-220
Number of pages8
JournalCell Stem Cell
Volume17
Issue number2
DOIs
Publication statusPublished - 2015 Aug 6

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Clustered Regularly Interspaced Short Palindromic Repeats
Induced Pluripotent Stem Cells
Factor VIII
Hemophilia A
Genes
High-Throughput Nucleotide Sequencing
Mutation
Inborn Genetic Diseases
Introns
Endothelial Cells
Genotype
Genome

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Medicine
  • Genetics

Cite this

Functional Correction of Large Factor VIII Gene Chromosomal Inversions in Hemophilia A Patient-Derived iPSCs Using CRISPR-Cas9. / Park, Chul Yong; Kim, Duk Hyoung; Son, Jeong Sang; Sung, Jin Jea; Lee, Jaehun; Bae, Sangsu; Kim, Jong-Hoon; Kim, Dong Wook; Kim, Jin Soo.

In: Cell Stem Cell, Vol. 17, No. 2, 1805, 06.08.2015, p. 213-220.

Research output: Contribution to journalArticle

Park, Chul Yong ; Kim, Duk Hyoung ; Son, Jeong Sang ; Sung, Jin Jea ; Lee, Jaehun ; Bae, Sangsu ; Kim, Jong-Hoon ; Kim, Dong Wook ; Kim, Jin Soo. / Functional Correction of Large Factor VIII Gene Chromosomal Inversions in Hemophilia A Patient-Derived iPSCs Using CRISPR-Cas9. In: Cell Stem Cell. 2015 ; Vol. 17, No. 2. pp. 213-220.
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