| RRC ID |
85340
|
| Author |
Motosugi N, Hasegawa K, Kurosaki N, Kawaguchi E, Izumi K, Iida Y, Higashiseto M, Yokoyama K, Sasaki A, Nakabayashi K, Fukuda A.
|
| Title |
Highly efficient XIST reactivation in female hPSC by transient dual inhibition of TP53 and DNA methylation during Cas9 mediated genome editing.
|
| Journal |
Stem Cell Res Ther
|
| Abstract |
The irreversible erosion of X-chromosome inactivation (XCI) due to repression of the long non-coding RNA XIST presents a major challenge for disease modeling and raises safety concerns for the clinical application of female human pluripotent stem cells (hPSCs) due to the aberrant overexpression of X-linked genes. While Cas9-mediated non-homologous end joining (NHEJ) targeting the XIST promoter can induce DNA demethylation and restore XCI by reactivating XIST, its efficiency remains low. Here, we introduce a highly efficient strategy for XIST reactivation by combining TP53 inhibition with suppression of DNA methylation maintenance during Cas9-mediated NHEJ. This dual-inhibition approach increased the proportion of XIST-positive hPSCs from ~ 5 to ~ 43.7%, providing a robust method for stabilizing XCI in female hPSCs for diverse applications.
|
| Volume |
16(1)
|
| Pages |
389
|
| Published |
2025-7-18
|
| DOI |
10.1186/s13287-025-04501-4
|
| PII |
10.1186/s13287-025-04501-4
|
| PMID |
40682144
|
| PMC |
PMC12275311
|
| MeSH |
CRISPR-Cas Systems* / genetics
DNA Methylation* / genetics
Female
Gene Editing* / methods
Humans
Pluripotent Stem Cells* / cytology
Pluripotent Stem Cells* / metabolism
RNA, Long Noncoding* / genetics
RNA, Long Noncoding* / metabolism
Tumor Suppressor Protein p53* / antagonists & inhibitors
Tumor Suppressor Protein p53* / genetics
Tumor Suppressor Protein p53* / metabolism
X Chromosome Inactivation / genetics
|
| IF |
5.116
|
| Resource |
| Human and Animal Cells |
HPS2478(HPS2478)
HPS2508(HPS2508)
HPS3131(HPS3131)
HPS3676(HPS3676)
HPS3681(HPS3681)
HPS3686(HPS3686)
HPS3692(HPS3692)
HPS3696(HPS3696)
HPS3810(HPS3810)
HPS4115(HPS4115) |
