A novel proteomics approach for identification of protein-RNA complex associated with specific gene loci using a human artificial chromosome (HAC)

为了恢复细胞中携带3.5kb lacO重复序列的CHO染色体，我们在引入LacI-GFP载体后，使用抗gfp抗体进行免疫沉淀。结果，抗gfp抗体恢复了大尺寸的CHO染色体。这些结果提示通过染色质免疫沉淀法恢复HAC的可能性。此外，我们的同事还开发了一种新的具有多整合酶重组位点的HAC载体（MI-HAC）。MI-HAC可以引入各种大基因或基因组位点。此外，我们制作了含有lacO重复序列和靶基因位点（Xist， LIT1/ kcnq10t1和CGG重复序列）的MI-HAC。

To recover the CHO chromosome that carry 3.5kb of the lacO repeat sequences in the cells, we carried out immunoprecipitation by using anti-GFP antibody after the introduction of the LacI-GFP vector. As a result, a large size of CHO chromosome was recovered by anti-GFP antibody. These results suggest that the possibility of recovery of the HAC by chromatin immunoprecipitation. In addition, our colleague has developed a new HAC vector (MI-HAC) with multi-integrase recombination sites. MI-HAC enables to introduce various large genes or genomic loci. Furthermore, we produced MI-HAC that contains the lacO repeat sequences and targeted gene loci (Xist, LIT1/KCNQ1OT1 and CGG repeat sequence).