Functional Characterization of IWS1 in Mouse Development

In this project, I led the functional in vivo characterization of IWS1, an RNA polymerase II-associated transcription elongation factor, by generating and analyzing a conditional knockout mouse model. IWS1 ablation resulted in pre-implantation embryonic lethality, establishing it as an essential gene for early development. Through comprehensive immunohistochemistry, Western blotting, and RT-qPCR, we showed that IWS1 is broadly expressed across adult tissues and highly enriched in oocytes and early embryos. Mechanistic studies suggested a critical role for IWS1 in coordinating transcription elongation with chromatin modifications and mRNA processing, particularly via interactions with the SETD2/H3K36me3 axis and RNAPII. These findings underscore the essential role of IWS1 in developmental epigenetic programming.

Genetic ablation of interacting with Spt6 (Iws1) causes early embryonic lethality

Arturo Orlacchio , Aaron E Stark, Claudia Foray, Foued Amari, Tyler Sheetz, Erika Reese, Anna Tessari, Krista La Perle, Dario Palmieri, Philip N Tsichlis, Vincenzo Coppola

AACR Annual Meeting 2018

Genetic ablation of interacting with Spt6 (Iws1) causes early embryonic lethality

Arturo Orlacchio , Aaron E Stark, Claudia Foray, Foued Amari, Tyler Sheetz, Erika Reese, Anna Tessari, Krista La Perle, Dario Palmieri, Philip N Tsichlis, Vincenzo Coppola

Abstract

IWS1 is an RNA-polymerase II (RNAPII)-associated transcription elongation factor whose biological functions are poorly characterized. To shed some light on the function of this protein at the organismal level, we performed a systematic tissue analysis of its expression and generated Iws1-deficient mice. A thorough immunohistochemical characterization shows that IWS1 protein is present in the nucleus of all cells in most of the examined tissues, with few notable exceptions. We also report that ablation of Iws1 consistently causes lethality at the pre-implantation stage with high expression of the gene in fertilized oocytes. In summary, we are providing evidence that Iws1 is expressed in all adult organs and it is an essential gene for mouse embryonic development.

PlosOne 2018