Directed evolution of a recombinase that excises the provirus of most HIV-1 primary isolates with high specificity
Directed evolution of a recombinase that excises the provirus of most HIV-1 primary isolates with high specificity
复制标题
DOI：
10.1038/nbt.3467
复制
发表时间：
2016-04-01
影响因子：
46.9
通讯作者：
Buchholz, Frank
中科院分区：
文献类型：
--
作者：
Karpinski, Janet;Hauber, Ilona;Buchholz, Frank
目前的联合抗逆转录病毒疗法（cART）可有效抑制人类HIV-1的繁殖，但该病毒在少数细胞中作为整合的原病毒库持续存在。为了产生一种能够根除受感染细胞中原病毒的抗病毒剂，我们采用了 145 个底物相关定向进化循环来进化出一种重组酶 (Brec1)，该酶可位点特异性识别大多数临床相关 HIV-1 毒株和亚型的长末端重复序列 (LTR) 中存在的 34 bp 序列。 Brec1 高效、精确且安全地从受感染细胞中去除整合的原病毒，并且对体外和体内临床 HIV-1 分离株有效，包括在用患者来源的细胞人源化的小鼠中。我们的数据表明 Brec1 具有作为 HIV-1 治疗药物的临床应用潜力。
Current combination antiretroviral therapies (cART) efficiently suppress HIV-1 reproduction in humans, but the virus persists as integrated proviral reservoirs in small numbers of cells. To generate an antiviral agent capable of eradicating the provirus from infected cells, we employed 145 cycles of substrate-linked directed evolution to evolve a recombinase (Brec1) that site-specifically recognizes a 34-bp sequence present in the long terminal repeats (LTRs) of the majority of the clinically relevant HIV-1 strains and subtypes. Brec1 efficiently, precisely and safely removes the integrated provirus from infected cells and is efficacious on clinical HIV-1 isolates in vitro and in vivo, including in mice humanized with patient-derived cells. Our data suggest that Brec1 has potential for clinical application as a curative HIV-1 therapy.