{"title":"Avian Retrovirus-Mediated Tumor-Specific Gene Knockout","authors":"Wei Wang, Bingning Dong, Feng Yang","doi":"10.1002/cpmb.54","DOIUrl":null,"url":null,"abstract":"<p>The RCAS (replication-competent avian sarcoma leukosis virus long-terminal repeat with splice acceptor)-TVA (tumor virus A) gene delivery system has been successfully used in modeling human cancers. Based on this, we have recently developed a novel RCI-Oncogene (RCAS-Cre-IRES-Oncogene) gene delivery system that can be used to efficiently manipulate gene expression in spontaneous tumors in vivo. We used this system for tumor gene knockout (TuKO) and demonstrated a crucial role of FGFR1 in driving mammary tumor metastasis. This versatile tumor gene modification system can also be adapted into different configurations to address different questions in appropriate mutant mouse hosts. Here we describe a protocol using the TuKO approach to knock out a gene of interest in tumors in appropriate hosts. © 2018 by John Wiley & Sons, Inc.</p>","PeriodicalId":10734,"journal":{"name":"Current Protocols in Molecular Biology","volume":"121 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2018-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpmb.54","citationCount":"2","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Molecular Biology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpmb.54","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 2
Abstract
The RCAS (replication-competent avian sarcoma leukosis virus long-terminal repeat with splice acceptor)-TVA (tumor virus A) gene delivery system has been successfully used in modeling human cancers. Based on this, we have recently developed a novel RCI-Oncogene (RCAS-Cre-IRES-Oncogene) gene delivery system that can be used to efficiently manipulate gene expression in spontaneous tumors in vivo. We used this system for tumor gene knockout (TuKO) and demonstrated a crucial role of FGFR1 in driving mammary tumor metastasis. This versatile tumor gene modification system can also be adapted into different configurations to address different questions in appropriate mutant mouse hosts. Here we describe a protocol using the TuKO approach to knock out a gene of interest in tumors in appropriate hosts. © 2018 by John Wiley & Sons, Inc.
禽逆转录病毒介导的肿瘤特异性基因敲除
RCAS(复制型禽肉瘤白血病病毒长末端重复与剪接受体)-TVA(肿瘤病毒A)基因传递系统已成功用于模拟人类癌症。基于此,我们最近开发了一种新的RCI-Oncogene (RCAS-Cre-IRES-Oncogene)基因传递系统,可用于有效地操纵体内自发性肿瘤中的基因表达。我们使用该系统进行肿瘤基因敲除(TuKO),并证明了FGFR1在驱动乳腺肿瘤转移中的关键作用。这种多功能肿瘤基因修饰系统也可以适应成不同的配置,以解决不同的问题,在适当的突变小鼠宿主。在这里,我们描述了一种使用TuKO方法在适当宿主中敲除肿瘤中感兴趣的基因的方案。©2018 by John Wiley &儿子,Inc。
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