Genome-wide CRISPR/Cas9 transcriptional activation screen identifies a histone acetyltransferase inhibitor complex as a regulator of HIV-1 integration.
Genome-wide CRISPR/Cas9 transcriptional activation screen identifies a histone acetyltransferase inhibitor complex as a regulator of HIV-1 integration.
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DOI：
10.1093/nar/gkac464
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发表时间：
2022-07-08
影响因子：
14.9
通讯作者：
Rana, Tariq M.
中科院分区：
文献类型：
--
作者：
Zhang, Qiong;Wang, Shaobo;Li, Wanyu;Yau, Edwin;Hui, Hui;Singh, Parmit Kumar;Achuthan, Vasudevan;Young Karris, Maile Ann;Engelman, Alan N.;Rana, Tariq M.
逆转录病毒人类免疫缺陷病毒-1（HIV-1）是艾滋病的病原体。虽然用抗逆转录病毒疗法治疗HIV/AIDS提供了对病毒血症的抑制，但整合的前病毒的潜伏储库妨碍了当前抗病毒疗法的治愈。了解宿主-病毒相互作用的机制可能阐明新的治疗策略。在这里，我们使用高复杂性的全基因组sgRNA文库进行了CRISPR/Cas9转录激活筛选，以鉴定抑制HIV-1感染人类CD 4 + T细胞的细胞因子。用CRISPR/Cas9 sgRNA文库转导MT4细胞，并用表达更昔洛韦敏感性胸苷激酶的nef缺陷型HIV-1 NL 4 -3感染，从而能够选择HIV-1抗性细胞用于分析富集的sgRNA。在验证筛选命中后，鉴定了HIV-1感染所必需的多种宿主因子，包括SET（SET核原癌基因）和ANP 32A（酸性核磷蛋白32A，PP 32A），它们一起形成组蛋白乙酰化酶抑制剂复合物。我们使用来自健康供体和HIV-1感染者的多种人类细胞系和外周血单核细胞（PBMCs），证明SET耗尽通过增加DNA整合而增加HIV-1的感染性，而不会显着改变整合位点。相反，SET过表达降低HIV-1整合和感染性。SET蛋白的表达显着减少，在PBMC中的HIV-1感染的个人和下调HIV-1感染的健康供体细胞在体外。值得注意的是，HIV-1诱导的SET下调可以通过抑制蛋白酶颗粒酶A来缓解。总而言之，我们已经在全基因组范围内确定了HIV-1感染的细胞抑制剂，这为宿主-病毒相互作用提供了新的见解，并可能为HIV-1治疗提供新的策略。
The retrovirus human immunodeficiency virus-1 (HIV-1) is the causative agent of AIDS. Although treatment of HIV/AIDS with antiretroviral therapy provides suppression of viremia, latent reservoirs of integrated proviruses preclude cure by current antiviral treatments. Understanding the mechanisms of host–viral interactions may elucidate new treatment strategies. Here, we performed a CRISPR/Cas9 transcriptional activation screen using a high-complexity, genome-wide sgRNA library to identify cellular factors that inhibit HIV-1 infection of human CD4+ T cells. MT4 cells were transduced with a CRISPR/Cas9 sgRNA library and infected with nef-deficient HIV-1NL4-3 expressing ganciclovir-sensitive thymidine kinase, thus enabling selection of HIV-1-resistant cells for analysis of enriched sgRNAs. After validation of screen hits, multiple host factors essential for HIV-1 infection were identified, including SET (SET nuclear proto-oncogene) and ANP32A (acidic nuclear phosphoprotein 32A, PP32A), which together form a histone acetylase inhibitor complex. Using multiple human cell lines and peripheral blood mononuclear cells (PBMCs) from healthy donors and HIV-1-infected individuals, we demonstrate that SET depletion increased HIV-1 infectivity by augmenting DNA integration without significantly changing sites of integration. Conversely, SET overexpression decreased HIV-1 integration and infectivity. SET protein expression was significantly reduced in PBMCs from HIV-1-infected individuals and was downregulated by HIV-1 infection of healthy donor cells in vitro. Notably, HIV-1-induced downregulation of SET could be alleviated by inhibition of the protease granzyme A. Altogether, we have identified cellular inhibitors of HIV-1 infection on a genome-wide scale, which affords new insight into host–virus interactions and may provide new strategies for HIV-1 treatment.
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DOI：
10.1093/bioinformatics/btq033
发表时间：
2010-03-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Quinlan AR;Hall IM
通讯作者：
Hall IM
影响因子：
64.8
作者：
Konermann S;Brigham MD;Trevino AE;Joung J;Abudayyeh OO;Barcena C;Hsu PD;Habib N;Gootenberg JS;Nishimasu H;Nureki O;Zhang F
通讯作者：
Zhang F
影响因子：
14.9
作者：
Desfarges S;San Filippo J;Fournier M;Calmels C;Caumont-Sarcos A;Litvak S;Sung P;Parissi V
通讯作者：
Parissi V
影响因子：
14.9
作者：
Bedwell GJ;Jang S;Li W;Singh PK;Engelman AN
通讯作者：
Engelman AN
影响因子：
1.2
作者：
Serrao, Erik;Cherepanov, Peter;Engelman, Alan N.
通讯作者：
Engelman, Alan N.