Physiological roles of Ewings Sarcoma gene product EWS

尤文氏肉瘤基因(EWS)属于Tet蛋白家族，包括TLS/FUS和TAFII15。这些蛋白质在C末端编码RNA结合域，在其N末端编码SYGOO基序的大量重复。基于与RNA聚合酶II和剪接因子的物理相互作用，Tet蛋白被认为在基本转录和剪接中发挥作用。然而，EWS和其他Tet蛋白在体内的作用尚未被证明。因此，本项目的目标是了解EWS的体内功能，并将该功能与尤文氏肉瘤和其他EWS融合肉瘤中发现的染色体易位联系起来。目的1.通过基因打靶的方法获得EWS基因敲除小鼠。我们已经从靶向小鼠ES细胞中产生了基因敲除小鼠。目的2.研究EWS缺陷对基因敲除小鼠的影响。目的3.在细胞水平上表征EWS缺陷，以确定EWS发挥作用的机制。从这项研究中获得的知识将有助于更好地了解EWS和其他Tet蛋白。这将进一步阐明EWS或TLS相关融合产品将如何导致肿瘤发生。

Ewing's sarcoma gene (EWS) belongs to a family of TET proteins, which include TLS/FUS and TAFII15. These proteins encode RNA binding domain at the C-terminal end and a large repeats of SYGOO motif at their N-terminus. Based on the physical interactions with RNA polymerase II and splicing factors, TET proteins are thought to play roles in basic transcription and splicing. However, in vivo role of EWS and other TET proteins has not been demonstrated. Thus, the goal of this project is to understand the in vivo function of EWS and to relate that function in the context of the chromosomal translocation found in Ewing's sarcoma and other EWS-fusion sarcomas. Aim 1. We will generate EWS knockout mouse by gene targeting approach. We have already generated the knockout mouse derived from the targeted mouse ES cells. Aim 2. To characterize the defects caused by EWS deficiency in the knockout mouse. Aim 3. To characterize the EWS deficiency at the cellular level to identify the mechanisms by which EWS functions. The knowledge gained from this study will help to better understand about EWS and other TET proteins in general. This will further elucidate how EWS or TLS related fusion products will lead to tumorigenesis.