Antisense Oligonucleotide-Mediated XIST Knockdown Modulates Endogenous Pluripotency-Associated Transcription Factor Expression in Porcine Female Somatic Cells

Abstract

Simple Summary Somatic cell reprogramming is an important technology for animal biotechnology, embryo engineering, and genetic improvement. However, the efficiency of reprogramming is limited by epigenetic barriers that restrict the activation of pluripotency-related genes. The long non-coding RNA XIST plays a central role in X-chromosome inactivation and contributes to epigenetic regulation during cell fate transitions. In this study, we investigated whether XIST knockdown could influence endogenous pluripotency-associated genes in IBRS-2 cells, a porcine female somatic cell line. Chemically modified antisense oligonucleotides (ASOs) were used to suppress XIST expression in IBRS-2 cells. XIST knockdown resulted in sustained increases in KLF4 and LIF expression, together with transient activation of NANOG and other pluripotency-associated genes. KLF4 protein was detectable by antibody staining in a subset of ASO-2-treated cells. These findings indicate that XIST suppression can facilitate the partial activation of endogenous pluripotency-associated genes and may provide a potential molecular strategy for modulating cell reprogramming-associated gene expression in pigs.