Targets and functions of the Kaposi's Sarcoma associated herpesvirus microRNAs

2010年10月，我将在西北大学担任独立教职，继续攻读 卡波西肉瘤相关疱疹病毒(KSHV)microRNAs(MiRNAs)的功能分析。MiRNAs是 -22个核苷酸长的非编码RNA，转录后抑制带有不完全匹配的mRNA 到miRNA，主要是在他们的3‘非编码区。KSHV表达12个前miRNAs的病毒miRNAs 在潜伏感染期间循环。在这个奖项的K99阶段，我全面确定了以下目标 所有使用新开发的PAR-CLIP技术的KSHV miRNAs，并表征了KSHV的作用 MJR-Ki在KSHV逃避P53介导的细胞周期停滞中的作用我对Roo阶段的研究目标是 该奖项旨在利用KSHVmiRNA目标的详细知识来研究 KSHV miRNAs对KSHV诱导的细胞周期进程和细胞凋亡的非调控作用。对这些的干扰 KSHV的途径被认为对这种病毒的致瘤特性至关重要。两国之间的相互作用 具有这些通路的KSHV miRNAs将被研究(A)基于这些通路的PAR-CLIP识别的靶 在细胞周期调节和诱导细胞凋亡中具有已知功能的miRNAs，(B)通过系统 异位表达单个或多个B细胞系或内皮细胞的功能询问 KSHV miRNAs，以及(C)通过对潜伏感染KSHV原发积液的系统功能询问 淋巴瘤(PEL)细胞或内皮细胞，其中单个或多个miRNAs的功能已被 用海绵堵住了。作为一名独立研究人员，我将通过定期的 与新成立的疱疹病毒学、miRNA生物学专家指导委员会的会议。 此外，我预计拟议的研究将产生足够的有趣数据来申请ROI 在独立阶段的早期授予，也极大地影响了我的长期研究计划。

In October 2010, I am moving to an independent faculty position at Northwestern University to pursue the functional analysis of Kaposi's saroma-associated herpesvirus (KSHV) microRNAs (miRNAs). MiRNAs are -22 nucleotide long non-coding RNAs that posttranscriptionally repress mRNAs bearing imperfect matches to the miRNA, primarily in their 3'UTR. KSHV expresses a number of viral miRNAs from 12 pre-miRNAstem loops during latent infection. During the K99 phase ofthis award, I have comprehensively identified targets of all KSHV miRNAs using the newly developed PAR-CLIP technology and characterized the role of KSHV mJR-KI in the evasion of p53-mediated cell cycle arrest by KSHV. My research goal for the ROO phase of this award is to use this detailed knowledge ofthe KSHVmiRNAtargets to study the contribution ofthe KSHV miRNAs to the de-regulation of cell cycle progression and apoptosis by KSHV. Interference with these pathways by KSHV is thought to be critical for the tumorigenic properties of this virus. The interplay of the KSHV miRNAs with these pathways will be studied (a) based on PAR-CLIP-identified targets of these miRNAs with known functions in cell cycle regulation and the induction of apoptosis, (b) through systematic functional interrogation of B-cell lines or endothelial cells which ectopically express individual or multiple KSHV miRNAs, and (c) through systematic functional interrogation of latently KSHV infected primary effusion lymphoma (PEL) cells or endothelial cells in which the function of individual or multiple miRNAs has been blocked using sponges. I will monitor my development as an independent researcher through regular meetings with a newly formed mentorship committee of experts in herpesvirology miRNA biology. Furthermore, I expect that the proposed research will yield sufficient interesting data to apply for an ROI grant early in the independent phase and also significantly shape my long term research program.