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

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) ge...

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Publicado en:Journal of Dental Research Vol. 99; no. 2; pp. 196 - 204
Autores principales: El Ayachi, I., Zou, X.-Y., Yan, X., Lou, Y., Huang, G.T.-J.
Formato: research Journal Article
Publicado: Sage Publications Inc. Feb2020
Acceso en línea:Ver este registro en EBSCOhost
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      dt: Feb2020
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      pub: Sage Publications Inc.
      place: Thousand Oaks, California
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        atl: Expression of Normal or Mutated X-Linked Transcripts in OFCD iPSCs.
      aug:
        au:
          El Ayachi, I.
          Zou, X.-Y.
          Yan, X.
          Lou, Y.
          Huang, G.T.-J.
        affil: Department of Bioscience Research, College of Dentistry, University of Tennessee Health Science Center, Memphis, TN, USA
      sug:
        subj:
          Eye Abnormalities
          Heart Septal Defects
          Tooth Root
          Stem Cells
          Mice
          Models, Biological
          Proteins
          Animal Studies
          Human
          United States
          Validation Studies
          Comparative Studies
          Evaluation Research
          Multicenter Studies
      ab: 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.
      pubtype: Academic Journal
      doctype:
        research
        Journal Article
      ougenre: Article
    language: English
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