Link Robert W, Nonnemacher Michael R, Wigdahl Brian, Dampier Will
1 School of Biomedical Engineering, Science and Health Systems, Drexel University, Philadelphia, Pennsylvania.
2 Department of Microbiology and Immunology, Drexel University College of Medicine, Philadelphia, Pennsylvania.
CRISPR J. 2018 Aug;1(4):294-302. doi: 10.1089/crispr.2018.0020.
Chronic human immunodeficiency virus type 1 (HIV-1) disease is characterized by the retention of provirus within latently infected cells. Anti-HIV-1 CRISPR-Cas9 gene editing is an attractive strategy to excise or inactivate the HIV-1 genome. Recent strategies have focused on designing gRNAs that target the long terminal repeat (LTR) because 5' and 3' LTR symmetry can facilitate proviral excision. However, the promiscuity of CRISPR-Cas9 gene editing system necessitates the investigation of potential off-target effects. Here, potential gRNAs designed from HIV-1 phylogenetic subtypes using the CRISPRseek tool were investigated. Across the LTR, it was found that certain regions show higher human homology than others. When using recommended cutoffs, 96.40% of gRNAs were predicted to have no high probability off-target effects. Given this observation, while high-probability off-target effects are a potential danger, they can be avoided with proper gRNA design.
慢性1型人类免疫缺陷病毒（HIV-1）疾病的特征是原病毒保留在潜伏感染的细胞内。抗HIV-1 CRISPR-Cas9基因编辑是切除或使HIV-1基因组失活的一种有吸引力的策略。最近的策略集中在设计靶向长末端重复序列（LTR）的引导RNA（gRNA），因为5'和3' LTR的对称性有助于前病毒的切除。然而，CRISPR-Cas9基因编辑系统的混杂性使得有必要研究潜在的脱靶效应。在此，对使用CRISPRseek工具从HIV-1系统发育亚型设计的潜在gRNA进行了研究。在整个LTR中，发现某些区域的人类同源性高于其他区域。使用推荐的临界值时，预计96.40%的gRNA没有高概率的脱靶效应。基于这一观察结果，虽然高概率脱靶效应是一个潜在危险，但通过适当的gRNA设计可以避免。
