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. 2019 Feb 28;9:3134. doi: 10.1038/s41598-019-40003-z

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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CRISPR-mediated enhancement of BST-2 expression leads to marked decrease in wild-type HIV-1 replication. (A) CD4-positive H9 cells with an intermediate level of BST-2 expression were transduced with lentiviral CRISPR activation vector expressing an empty sgRNA (H9-sgCtrl) or an sgRNA targeting BST-2 promoter (H9-sgBST2), the latter of which were cloned. Cells were analyzed by real-time RT-PCR for BST-2 expression. Representative data from three independent experiments are shown as a fold difference in BST-2 copies compared with those in HeLa cells (mean ± s.d., n = 3 technical replicates). (B) Cell proliferation of CRISPR-transduced H9 cells (H9-sgCtrl, closed circles; H9-sgBST2, open circles). The assay was started with 20,000 cells. Data shown are representative of three independent experiments. (C) Virus replication in H9-sgCtrl (closed symbols) or H9-sgBST2 (open symbols). Cells (1 × 10⁵) were infected with 0.1 ng of p24 antigen of either wild-type (squares) or Vpu-defective (triangles) NL4-3 viruses. Supernatants were harvested at two-to-three-day intervals, and virus replication was monitored by p24 ELISA. Data shown are representative of four independent experiments.