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To overcome these drawbacks, we have developed a single lentiviral vector that can concurrently deplete endogenous gene expression while expressing an epitope-tagged siRNA-resistant target gene in the same cell. To demonstrate the functional utility of this system, we performed RNAi-depleted \u03b1-actinin-1 (\u03b1-ACTN1) expression in human T cells. \u03b1-ACTN1 RNAi resulted in inhibited chemotaxis to SDF-1\u03b1, but it can be completely rescued by concurrent expression of RNAi-resistant \u03b1-ACTN1 (rr-\u03b1-ACTN1) in the same cell. The presence of a GFP tag on rr-\u03b1-ACTN1 allowed for detection of appropriate subcellular localization of rr-\u03b1-ACTN1. This system provides not only an internal control for RNAi off-target effects, but also the potential tool for rapid structure-function analyses and gene therapy.<\/jats:p>","DOI":"10.1016\/s1672-0229(10)60025-3","type":"journal-article","created":{"date-parts":[[2011,3,8]],"date-time":"2011-03-08T04:31:43Z","timestamp":1299558703000},"page":"238-245","source":"Crossref","is-referenced-by-count":39,"title":["A Multifunctional Lentiviral-Based Gene Knockdown with Concurrent Rescue that Controls for Off-Target Effects of RNAi"],"prefix":"10.1093","volume":"8","author":[{"given":"Yunfeng","family":"Feng","sequence":"first","affiliation":[{"name":"Departments of Medicine and Cell Biology, Washington University , St. Louis, MO 63110 , USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Linghu","family":"Nie","sequence":"additional","affiliation":[{"name":"Key Laboratory of Genome Sciences and Information, Cancer Biology and Genetics Group, Beijing Institute of Genomics, Chinese Academy of Sciences , Beijing 100029 , China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Meghna Das","family":"Thakur","sequence":"additional","affiliation":[{"name":"Departments of Medicine and Cell Biology, Washington University , St. Louis, MO 63110 , USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qin","family":"Su","sequence":"additional","affiliation":[{"name":"Immunocore Limited , Abingdon, Oxon OX14 4RX , UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhenfen","family":"Chi","sequence":"additional","affiliation":[{"name":"Key Laboratory of Genome Sciences and Information, Cancer Biology and Genetics Group, Beijing Institute of Genomics, Chinese Academy of Sciences , Beijing 100029 , China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yongliang","family":"Zhao","sequence":"additional","affiliation":[{"name":"Key Laboratory of Genome Sciences and Information, Cancer Biology and Genetics Group, Beijing Institute of Genomics, Chinese Academy of Sciences , Beijing 100029 , China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gregory D.","family":"Longmore","sequence":"additional","affiliation":[{"name":"Departments of Medicine and Cell Biology, Washington University , St. Louis, MO 63110 , USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2011,3,5]]},"reference":[{"key":"2024051008274111600_bib1","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1016\/S1535-6108(02)00122-8","article-title":"Stable suppression of tumorigenicity by virus-mediated RNA interference","volume":"2","author":"Brummelkamp","year":"2002","journal-title":"Cancer Cell"},{"key":"2024051008274111600_bib2","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1073\/pnas.232688199","article-title":"Inhibiting HIV-1 infection in human T cells by lentiviral-mediated delivery of small interfering RNA against CCR5","volume":"100","author":"Qin","year":"2003","journal-title":"Proc. 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