日前，朱煥章課題組利用CRISPR/CAS9技術在體外靶向幹預HIV潛伏又取得進展，為艾滋的功能治愈邁出了關鍵一步。該文發表在國際基因治療領域著名雜誌Mol Ther.（醫學1區）在線發表（http://www.ncbi.nlm.nih.gov/pubmed/26775808），相關工作亦已申請了中國專利。
目前，國際上已有近30種抗逆轉錄病毒藥物應用於艾滋病的臨床治療，然而，現有這些藥物隻能抑製病毒複製,不能清除艾滋病病毒（HIV）感染, 其原因主要在於HIV可潛伏於少數免疫細胞，以此逃脫了機體免疫係統和抗病毒藥物攻擊。因此，研發能靶向幹預HIV潛伏的新型治療技術，已成為目前國際上艾滋病治愈研究領域發展趨勢和研究的焦點。
基因編輯技術已有多個臨床試驗方案已被美國FDA批準應用到病人的抗腫瘤或艾滋病的基因治療研究。該文研究者提出了利用CRISPR/Cas9技術靶向激活潛伏HIV前病毒的“引蛇出洞”的策略。研究人員設計並篩選出了能特異靶向多數HIV亞型基因保守區的dCas9-SunTag-VP64 係統，在多個HIV潛伏細胞係上證實了dCas9-SunTag-VP64 係統能特異靶向激活HIV-1前病毒，提示了該方法有可能成為一個可選擇的抗HIV潛伏治療手段。
近幾年來，朱煥章課題組在艾滋病基因治療新型策略研究方麵取得了階段性的研究進展。2013年，首次提出並在體外證實利用基因編輯技術靶向切除整合HIV前病毒的“斬草除根”的AIDS/HIV治療策略的可行性（Nucleic Acids Res，2013;41(16):7771-82）；2014年，又獲得了能特異靶向HIV調控區的鋅指轉錄因子（ＺＦＰ）及轉錄樣因子（ＴＡＬＥ）融合基因，並在潛伏細胞模型上證實了這些融合蛋白可靶向激活潛伏ＨＩＶ表達（Gene Therapy ，2014，21，90-95；AIDS Res Hum Retroviruses，2015，31:98-106）。
Specific Reactivation of Latent HIV-1 by dCas9-SunTag-VP64-mediated Guide RNA Targeting the HIV-1 Promoter
Abstract：HIV-1 escapes antiretroviral agents by integrating into the host DNA and forming a latent transcriptionally silent HIV-1 provirus. Transcriptional activation is prerequisite for reactivation and the eradication of latent HIV-1 proviruses. dCas9-SunTag-VP64 transcriptional system has been reported that it can robustly activate the expression of an endogenous gene using a single guide RNA (sgRNA). Here, we systematically investigated the potential of dCas9-SunTag-VP64 with the designed sgRNAs for reactivating latent HIV-1. We found dCas9-SunTag-VP64 with sgRNA 4 or sgRNA 5 targeted from -164 to -146 or -124 to -106 bp upstream of the transcription start sites of HIV-1 could induce high expression of luciferase reporter gene after screening of sgRNAs targeting different regions of the HIV-1 promoter. Further, we confirmed that dCas9-SunTag-VP64 with sgRNA 4 or sgRNA 5 can effectively reactivate latent HIV-1 transcription in several latently infected human T-cell lines. Moreover, we confirmed that the reactivation of latent HIV-1 by dCas9-SunTag-VP64 with the designed sgRNA occurred through specific binding to the HIV-1 LTR promoter without genotoxicity and global T-cell activation. Taken together, our data demonstrated dCas9-SunTag-VP64 system can effectively and specifically reactivate latent HIV-1 transcription, suggesting that this strategy could offer a novel approach to anti-HIV-1 latency. （Mol Ther. 2016 Mar;24(3):508-21. doi: 10.1038/mt.2016.7. Epub 2016 Jan 18.）