LSm14A 参与抗病毒天然免疫反应的功能和分子机制研究

细胞受病毒感染后，胞内的模式识别受体识别病毒核酸，产生抗病毒天然免疫反应。LSm14A是新鉴定的病毒核酸受体，能够识别病毒DNA和RNA，在细胞抗病毒天然免疫中起重要作用。为深入研究LSm14A 参与天然免疫的功能和分子机制，我们构建了LSm14A的基因敲入（KI）细胞系和基因敲除（KO）小鼠。利用LSm14A KI 细胞系，通过亲和纯化和质谱分析，我们可以筛选与LSm14A相互作用蛋白。此外，我们将利用不同类型的模式病毒感染不同类型的LSm14A-/-和WT小鼠细胞（成纤维细胞、巨噬细胞等），研究LSm14A识别病毒核酸的特异性；我们还将利用LSm14A-/-小鼠深入研究 LSm14A 在机体抗病毒天然免疫反应中的功能。该项目的成功完成将为了解天然免疫对病毒的精密识别和调控机制提供重要依据。

The innate immune system recognizes viral nucleic acids by pattern recognition receptors which in turn mediate type I IFN induction and innate antiviral response. LSm14A is a newly identified viral nucleic acid sensor, which detects both viral RNA and DNA and mediates the expressions of antiviral genes in the early phage of viral infection. In order to understand the function and molecular mechanisms of LSm14A mediated antiviral response, we constructed HCT116 cell lines in which a Flag tag has been knocked in to the 3' of the endogenous LSm14A gene. We have also constructed LSm14A knock-out mice. To identify LSm14A interacting proteins, we have used HCT116-LSm14A-Flag cells for affinity purification with anti-Flag antibody. To determine the nucleic acid binding specificity, we will infect LSm14A-/- and WT cells with various DNA and RNA viruses. Moreover, on an animal level, we will use LSm14A-/- and WT mice to investigate the physiological function of LSm14A in innate immnune response against various viruses. This project will enrich our understanding of the recognition and regulatory mechanisms of antiviral innate immunity.