Dataset Information


CRISPR/Cas9-mediated modulation of splicing efficiency reveals short splicing isoform of Xist RNA is sufficient to induce X-chromosome inactivation.

ABSTRACT: Alternative splicing of mRNA precursors results in multiple protein variants from a single gene and is critical for diverse cellular processes and development. Xist encodes a long noncoding RNA which is a central player to induce X-chromosome inactivation in female mammals and has two major splicing variants: long and short isoforms of Xist RNA. Although a differentiation-specific and a female-specific expression of Xist isoforms have been reported, the functional role of each Xist RNA isoform is largely unexplored. Using CRISPR/Cas9-mediated targeted modification of the 5' splice site in Xist intron 7, we create mutant female ES cell lines which dominantly express the long- or short-splicing isoform of Xist RNA from the inactive X-chromosome (Xi) upon differentiation. Successful execution of CRISPR/Cas-based splicing modulation indicates that our CRISPR/Cas-based targeted modification of splicing sites is a useful approach to study specific isoforms of a transcript generated by alternative splicing. Upon differentiation of splicing-mutant Xist female ES cells, we find that both long and short Xist isoforms can induce X-chromosome inactivation normally during ES cell differentiation, suggesting that the short splicing isoform of Xist RNA is sufficient to induce X-chromosome inactivation.


PROVIDER: S-EPMC5861412 | BioStudies | 2018-01-01

REPOSITORIES: biostudies

Similar Datasets

2019-01-01 | S-EPMC6379716 | BioStudies
2006-01-01 | S-EPMC1464350 | BioStudies
1000-01-01 | S-EPMC4278889 | BioStudies
2009-01-01 | S-EPMC2720191 | BioStudies
2007-01-01 | S-EPMC2121114 | BioStudies
2017-01-01 | S-EPMC5521851 | BioStudies
1000-01-01 | S-EPMC2546918 | BioStudies
2016-01-01 | S-EPMC5064214 | BioStudies
2013-05-30 | E-GEOD-46918 | BioStudies
1000-01-01 | S-EPMC3029249 | BioStudies