采用Cas9-Spinach2 RNA 荧光标记系统研究IRAIN 在AML干细胞中的功能和机制

白血病干细胞（LSC）是导致急性髓系白血病(AML)发生、化疗抵抗最终难治复发的根源之一。胰岛素样生长因子1受体(IGF1R) 与白血病生长化疗耐药密切相关，但IGF1R在LSC中异常表达的调控机制不清楚。. 申请人在国际上首次发现IGF1R基因内一条长链非编码RNA-IRAIN，开创一种研究染色质空间结构的新方法-逆转录结合染色质构象捕获法（RAT），第一作者文章发表在《Nucleic Acids Research》IF=9.112），IRAIN被NCBI收录（Gene ID: 104472848）；前期结果证实在LSC中IRAIN可能作为抑癌基因调控白血病的发生。. 本项目拟在前期工作基础上，结合CRISPR-Cas9基因编辑及Spinach2 RNA荧光定位分析，进一步阐明IRAIN 调控急性髓系白血病干细胞生长的功能和表观遗传学机制，为靶向治疗及新药开发提供理论依据。

Leukemia stem cells (LSC) induces the progresses and drug resistance of acute myeloid leukemia. Insulin-like growth factor 1 receptor（IGF1R) is often dysregulated in leukemia，particularly in LSC, but the underlying mechanism still remains to be defined. . Previously, we for the first time discovered a novel antisense long noncoding RNA, IRAIN, in the IGF1R gene locus. Using a reverse transcription-associated trap（RAT）method established in our lab, we found that IRAIN participated in the regulation of IGF1R by forming lncRNA/chromatin conformation looping. The data have been published on《Nucleic Acids Research》，and IRAIN has been recorded on NCBI（Gene ID: 104472848）. Importantly, we found that IRAIN may function as a tumor suppressor in LSC.. In this project, we propose to study the function and mechanism of IRAIN in LSC by using CRISPR Cas9-Spinach2 fluorescence-labeling system and RAT methods. We hope that the data from this project will provide a mechanistic rationale for designing a new generation anti-leukemia drug that targets LSC by harnessing the IRAIN epigenetic machinery.