肺癌是癌症死亡的首要因素，我国由于空气污染和控烟不力，每年约有70万新增患者。研究显示，在占肺癌40%的肺腺癌中，有超过1000个基因展现一定频率的突变或表观遗传改变，如何鉴别驱动突变是困扰精准医疗的关键问题。本课题将围绕这一核心困扰，拟在K-rasG12D和Cas9/dCas9条件敲除小鼠中建立基于CRISPR-Cas9基因敲除/过表达的遗传学筛选和功能研究的肺腺癌模型。在此基础上，联用激光微切割技术，在该敲除模型中进行表观驱动基因的筛选和功能验证，并在此模型中引入近红外荧光iRFP和透明化技术在肺的原位分析肿瘤的表型和微环境。前期的敲除预实验显示，这个系统支持一定规模的遗传学筛选以及候选基因的功能分析。该模型的建成既能极大地缩短肺癌的在体遗传学研究周期从3-5年至1-2年，又能通过筛选找到新的抑癌基因，可基本克服小鼠遗传学资源缺乏问题。

Lung cancer is the leading cause of cancer death. China has about 700,000 new patients each year due to air pollution and poor control of tobacco. Studies have shown that more than 1,000 genes exhibit mutations or epigenetic changes in lung adenocarcinoma which occupies 40% of lung cancer. How to identify driver mutations is a key issue for precision medicine. This project will focus on this question and propose to establish a lung adenocarcinoma model based on CRISPR-Cas9 knockout/overexpression technology in K-rasG12D and Cas9/dCas9 conditional knockout mice. Based on this model, through combining laser capture microdissection technology, the potential driven genes will be screened and functionally verified in the knock-out model. We will also develop iRFP and tissue clearing technology by uDisco to examine tumor phenotype in situ. The preliminary data show that this system supports sub genome wide genetic screening and functional analysis of candidate genes. The completion of this project can shorten the in vivo lung cancer genetic research from 3-5 years to 1-2 years, find new tumor suppressor genes, and overcome the bottleneck of mouse genetic resources limitation.