Expression of Normal or Mutated X-Linked BCOR 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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Bibliographic Details
Published in:Journal of dental research 2020-02, Vol.99 (2), p.196-203
Main Authors: El Ayachi, I., Zou, X.-Y., Yan, X., Lou, Y., Huang, G.T.-J.
Format: Article
Language:English
Subjects:
Animals
Bcl-6 protein
Clonal deletion
Cloning
Dentistry
Developmental disabilities
Disease Models, Animal
Gene deletion
Heart Septal Defects
Humans
Induced Pluripotent Stem Cells
Mice
Microphthalmos
Mutation
Pluripotency
Proto-Oncogene Proteins - genetics
Repressor Proteins - genetics
Research Reports
Stem cells
Tooth Apex - cytology
Transgenes
United States
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Summary: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.
ISSN:0022-0345
1544-0591
DOI:10.1177/0022034519890323