当前位置: X-MOL 学术Retrovirology › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
CCR5 editing by Staphylococcus aureus Cas9 in human primary CD4+ T cells and hematopoietic stem/progenitor cells promotes HIV-1 resistance and CD4+ T cell enrichment in humanized mice
Retrovirology ( IF 3.3 ) Pub Date : 2019-06-11 , DOI: 10.1186/s12977-019-0477-y
Qiaoqiao Xiao , Shuliang Chen , Qiankun Wang , Zhepeng Liu , Shuai Liu , Huan Deng , Wei Hou , Dongcheng Wu , Yong Xiong , Jiafu Li , Deyin Guo

BackgroundThe chemokine receptor CCR5, which belongs to the superfamily of G protein-coupled receptors, is the major co-receptor for HIV-1 entry. Individuals with a homozygous CCR5Δ32 mutation have a long lasting and increased resistance to HIV-1 infection. Therefore, CCR5 represents an optimal target for HIV-1/AIDS gene therapy. The CRISPR/Cas9 system has been developed as one of the most efficacious gene editing tools in mammalian cells and the small-sized version from Staphylococcus aureus (SaCas9) has an advantage of easier delivery compared to the most commonly used version from Streptococcus pyogenes Cas9 (SpCas9).ResultsHere, we demonstrated that CCR5 could be specifically and efficiently edited by CRISPR/SaCas9 together with two sgRNAs, which were identified through a screening of 13 sgRNAs. Disruption of CCR5 expression by lentiviral vector-mediated CRISPR/SaCas9 led to increased resistance against HIV-1 infection in human primary CD4+ T cells. Moreover, humanized mice engrafted with CCR5-disrupted CD4+ T cells showed selective survival and enrichment when challenged with CCR5 (R5)-tropic HIV-1 in comparison to mock-treated CD4+ T cells. We also observed CCR5 could be targeted by CRISPR/SaCas9 in human CD34+ hematopoietic stem/progenitor cells without obvious differentiation deficiencies.ConclusionsThis work provides an alternative approach to disrupt human CCR5 by CRISPR/SaCas9 for a potential gene therapy strategy against HIV-1/AIDS.

中文翻译:

金黄色葡萄球菌 Cas9 在人原代 CD4+ T 细胞和造血干/祖细胞中编辑 CCR5 可促进人源化小鼠的 HIV-1 抗性和 CD4+ T 细胞富集

背景趋化因子受体CCR5属于G蛋白偶联受体超家族,是HIV-1进入的主要共同受体。具有纯合 CCR5Δ32 突变的个体对 HIV-1 感染具有持久且增强的抵抗力。因此,CCR5 代表了 HIV-1/AIDS 基因治疗的最佳靶点。CRISPR/Cas9 系统已被开发为哺乳动物细胞中最有效的基因编辑工具之一,与来自化脓性链球菌 Cas9 的最常用版本相比,来自金黄色葡萄球菌 (SaCas9) 的小型版本具有更容易递送的优势。 SpCas9)。结果在这里,我们证明了 CCR5 可以被 CRISPR/SaCas9 和两个 sgRNA 特异性有效地编辑,这两个 sgRNA 通过 13 个 sgRNA 的筛选确定。慢病毒载体介导的 CRISPR/SaCas9 干扰 CCR5 表达导致人类原代 CD4+ T 细胞对 HIV-1 感染的抵抗力增加。此外,与模拟处理的 CD4+ T 细胞相比,植入 CCR5 破坏的 CD4+ T 细胞的人源化小鼠在用 CCR5 (R5) 嗜性 HIV-1 攻击时表现出选择性存活和富集。我们还观察到 CRISPR/SaCas9 可以在没有明显分化缺陷的人类 CD34+ 造血干/祖细胞中靶向 CCR5。结论这项工作提供了一种替代方法,通过 CRISPR/SaCas9 破坏人类 CCR5,从而获得针对 HIV-1/AIDS 的潜在基因治疗策略. 与模拟处理的 CD4+ T 细胞相比,移植了 CCR5 破坏的 CD4+ T 细胞的人源化小鼠在用 CCR5 (R5) 嗜性 HIV-1 攻击时表现出选择性存活和富集。我们还观察到 CRISPR/SaCas9 可以在没有明显分化缺陷的人类 CD34+ 造血干/祖细胞中靶向 CCR5。结论这项工作提供了一种替代方法,通过 CRISPR/SaCas9 破坏人类 CCR5,从而获得针对 HIV-1/AIDS 的潜在基因治疗策略. 与模拟处理的 CD4+ T 细胞相比,移植了 CCR5 破坏的 CD4+ T 细胞的人源化小鼠在用 CCR5 (R5) 嗜性 HIV-1 攻击时表现出选择性存活和富集。我们还观察到 CRISPR/SaCas9 可以在没有明显分化缺陷的人类 CD34+ 造血干/祖细胞中靶向 CCR5。结论这项工作提供了一种替代方法,通过 CRISPR/SaCas9 破坏人类 CCR5,从而获得针对 HIV-1/AIDS 的潜在基因治疗策略.
更新日期:2019-06-11
down
wechat
bug