Analysis for chromosomal regulation of HIV gene expression by proviral labeling

本研究的目的是了解染色体对HIV表达的调控。我们计划利用LACO/I系统了解HIV前病毒的空间和染色体调控。要应用LACO/I系统，必须将Lac O重复序列插入体内的HIV前病毒。因此，我们首先探索了CRISPR/Cas9系统编辑HIV-1基因组的潜力。我们发现CRISPR/Cas9系统有效地切割和突变了T细胞中的HIV前病毒，导致了表达的干扰。此外，我们还利用CRISPR/Cas9系统建立了一种通过HDR修复途径诱导HIV前病毒精确突变的新方法。提示这些技术能够将LACO重复序列插入到HIV前病毒中，并有可能发现染色体对HIV基因表达的调节。

Aim of this study is to understand the chromosomal regulation for HIV expression. We planned to understand the spatial and chromosomal regulation of HIV provirus by using LacO/I system. To apply LacO/I system, Lac O repeat sequence must be inserted into HIV provirus in vivo. Therefore, we explored the potential of CRISPR/Cas9 system to edit HIV-1 genome as the first step. We found that CRISPR/Cas9 system efficiently cleaved and mutated HIV provirus in T cells, resulting in disturbing the expression. Furthermore, we established a new method that induces precise mutation in HIV provirus through HDR repair pathway using CRISPR/Cas9 system. It is suggested that these technologies enable to insert LacO repeat sequence into HIV provirus and have a potential to discover chromosomal regulation of HIV gene expression.