RRC ID 53893
Author Tran THY, Fukuda A, Aizawa S, Bui PL, Hayashi Y, Nishimura K, Hisatake K.
Title Live cell imaging of X chromosome reactivation during somatic cell reprogramming.
Journal Biochem Biophys Rep
Abstract Generation of induced pluripotent stem cells (iPSCs) with naive pluripotency is important for their applications in regenerative medicine. In female iPSCs, acquisition of naive pluripotency is coupled to X chromosome reactivation (XCR) during somatic cell reprogramming, and live cell monitoring of XCR is potentially useful for analyzing how iPSCs acquire naive pluripotency. Here we generated female mouse embryonic stem cells (ESCs) that carry the enhanced green fluorescent protein (EGFP) and humanized Kusabira-Orange (hKO) genes inserted into an intergenic site near either the Syap1 or Taf1 gene on both X chromosomes. The ESC clones, which initially expressed both EGFP and hKO, inactivated one of the fluorescent protein genes upon differentiation, indicating that the EGFP and hKO genes are subject to X chromosome inactivation (XCI). When the derived somatic cells carrying the EGFP gene on the inactive X chromosome (Xi) were reprogrammed into iPSCs, the EGFP gene on the Xi was reactivated when pluripotency marker genes were induced. Thus, the fluorescent protein genes inserted into an intergenic locus on both X chromosomes enable live cell monitoring of XCI during ESC differentiation and XCR during reprogramming. This is the first study that succeeded live cell imaging of XCR during reprogramming.
Volume 15
Pages 86-92
Published 2018-9-1
DOI 10.1016/j.bbrep.2018.07.007
PII S2405-5808(18)30098-0
PMID 30094351
PMC PMC6073053
DNA material B6N Mouse BAC clone (RDB07573) B6Ng01-177J10 B6Ng01-316J16
Human and Animal Cells BRC6(AES0010)