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Expression of Normal or Mutated X-Linked BCOR Transcripts in OFCD iPSCs.

Authors
  • El Ayachi, I1
  • Zou, X-Y2, 3
  • Yan, X3
  • Lou, Y1, 3
  • Huang, G T-J1, 3
  • 1 Department of Bioscience Research, College of Dentistry, University of Tennessee Health Science Center, Memphis, TN, USA.
  • 2 Department of Cariology, Endodontology and Operative Dentistry, School and Hospital of Stomatology, Peking University, Beijing, China. , (China)
  • 3 Department of Endodontics, Henry M. Goldman School of Dental Medicine, Boston University, Boston, MA, USA.
Type
Published Article
Journal
Journal of dental research
Publication Date
Feb 01, 2020
Volume
99
Issue
2
Pages
196–203
Identifiers
DOI: 10.1177/0022034519890323
PMID: 31775564
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

Reprogramming diseased cells with mutated genes into induced pluripotent stem cells (iPSCs) can allow studies of disease mechanism and correct the mutation. Oculofaciocardiodental (OFCD) syndrome is a developmental disorder caused by heterozygous mutations in the X-linked BCL-6 corepressor (BCOR) gene. In this present study, we aimed to reprogram stem cells from a tooth apical papilla (SCAP) of a patient with OFCD, termed SCAP-O, into iPSCs. The SCAP-O carry a copy of the BCOR gene having 1 nucleotide deletion in 1 of the alleles, therefore harboring a mixture of cells expressing either normal (SCAP-OBCOR-WT) or mutated (SCAP-OBCOR-mut) BCOR transcripts. We subcloned SCAP-O and separated SCAP-OBCOR-WT and SCAP-OBCOR-mut as verified by sequencing. The selected subclone SCAP-OBCOR-mut expressed only the mutated BCOR transcripts and remained in such condition after multiple passages. We reprogrammed SCAP-O and subclone SCAP-OBCOR-mut into transgene-free iPSCs using an excisable lentiviral vector system (hSTEMCCA-loxP) carrying 4 reprogramming factors in a single cassette, followed by removal of transgenes via Cre-mediated excision. We found that after reprogramming SCAP-O or subclone SCAP-OBCOR-mut into iPSCs, some of the iPSC clones expressed either solely the normal BCOR-WT or BCOR-mut transcripts, while other clones expressed both BCOR-WT and BCOR-mut transcripts. This is our first step toward establishing OFCD study models by generating isogenic control BCOR-WT iPSCs versus BCOR-mut iPSCs.

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