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Characterization of human-induced pluripotent stem cells carrying homozygous RB1 gene deletion.
- Source :
-
Genes to cells : devoted to molecular & cellular mechanisms [Genes Cells] 2020 Jul; Vol. 25 (7), pp. 510-517. Date of Electronic Publication: 2020 Apr 28. - Publication Year :
- 2020
-
Abstract
- Retinoblastoma is an infant cancer that results from loss of RB1 expression in both alleles. The RB1 gene was the first reported cancer suppressor gene; however, the mechanism by which RB1 loss causes cancer in the retina has not yet been clarified. Human-induced pluripotent stem cells (iPSCs) provide an ideal tool for mechanistic research regarding retinoblastoma. However, because RB1 is a tumor suppressor, loss of both alleles of RB1 in human iPS cells may affect the phenotype of the cells. To examine this possibility, we established human iPSCs with deletions in both alleles of RB1 by CRISPR/Cas9 technique to characterize the associated phenotype. We first examined the expression of RB1 transcripts by RT-qPCR, and RB1 transcripts were expressed in immature hiPSCs and then the expression levels of RB1 transcripts consistently increased during retinal organoid differentiation in human iPSCs. Expression levels of immature markers including SSEA4, OCT3/4 and NANOG were indistinguishable between control iPSCs and RB1 knockout iPSCs. Proliferative activity was also unaffected by homozygous RB1 deletion. Taken together, we showed that homozygous deletion of RB1 did not affect the maturation and proliferation statuses of human iPSCs.<br /> (© 2020 Molecular Biology Society of Japan and John Wiley & Sons Australia, Ltd.)
- Subjects :
- CRISPR-Cas Systems
Gene Deletion
Humans
Induced Pluripotent Stem Cells cytology
Nanog Homeobox Protein metabolism
Octamer Transcription Factor-3 metabolism
Phenotype
Retina growth & development
Retinal Neoplasms genetics
Retinal Neoplasms metabolism
Retinoblastoma genetics
Retinoblastoma metabolism
Retinoblastoma Binding Proteins metabolism
Stage-Specific Embryonic Antigens metabolism
Ubiquitin-Protein Ligases metabolism
Cell Differentiation genetics
Cell Proliferation genetics
Induced Pluripotent Stem Cells metabolism
Organoids metabolism
Retina metabolism
Retinoblastoma Binding Proteins genetics
Ubiquitin-Protein Ligases genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1365-2443
- Volume :
- 25
- Issue :
- 7
- Database :
- MEDLINE
- Journal :
- Genes to cells : devoted to molecular & cellular mechanisms
- Publication Type :
- Academic Journal
- Accession number :
- 32277725
- Full Text :
- https://doi.org/10.1111/gtc.12771