Yone Hideyuki, Kawashima Yuri, Hirai Hayato, Oda Arisa H, Sato Moritoshi, Kono Hiromitsu, Ohta Kunihiro
Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Komaba 3-8-1, Meguro-ku, Tokyo 153-8902, Japan.
Kanagawa Institute of Industrial Science and Technology (KISTEC), 3-2-1 Sakado, Takatsu-ku, Kawasaki, Kanagawa 213-0012, Japan.
Nucleic Acids Res. 2025 Apr 10;53(7). doi: 10.1093/nar/gkaf206.
TAQing technologies are based on the restriction enzyme-induced DNA double-strand break (DSB) formation in living cells, which results in large-scale genomic rearrangements and phenotypic alterations. Originally, the TAQing system requires heat treatments to activate restriction enzymes, which sometimes leads to cell toxicity or stress responses. Here, we developed a blue-light-controlled MagTAQing system, which induces DSBs exclusively upon blue-light exposure by assembling the split restriction enzymes via Magnet modules. Application of MagTAQing to mitotic budding yeast cells successfully triggered various genomic rearrangements upon blue-light exposure. Since this technology enables the conditional induction of genomic rearrangements in specific cells or tissues, we employed MagTAQing on meiotic yeast cells lacking the recombinase Spo11 to induce artificial DSBs. Consequently, Spo11-independent meiotic DSBs resulted in aneuploidies and nonallelic homologous recombinations between repetitive sequences such as ribosomal DNA and retrotransposons. These results suggest a pivotal role of Spo11-induced recombination in preventing chromosomal abnormality.
TAQing技术基于限制酶在活细胞中诱导DNA双链断裂（DSB）的形成，这会导致大规模的基因组重排和表型改变。最初，TAQing系统需要热处理来激活限制酶，这有时会导致细胞毒性或应激反应。在这里，我们开发了一种蓝光控制的MagTAQing系统，该系统通过磁体模块组装切割后的限制酶，仅在蓝光照射下诱导DSB。将MagTAQing应用于有丝分裂出芽酵母细胞，在蓝光照射下成功触发了各种基因组重排。由于这项技术能够在特定细胞或组织中条件性诱导基因组重排，我们在缺乏重组酶Spo11的减数分裂酵母细胞上使用MagTAQing来诱导人工DSB。因此，不依赖Spo11的减数分裂DSB导致了非整倍体以及核糖体DNA和逆转座子等重复序列之间的非等位同源重组。这些结果表明Spo11诱导的重组在预防染色体异常中起关键作用。
