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An ultra-efficient and convenient method for meiotic DNA break profiling in multiple organisms
Collaborating research groups in China have developed a novel technique, called DNA End tailing and sequencing (DEtail-seq), for meiotic DNA double-strand break (DSB) profiling.
In eukaryotes, meiosis is a fundamental process required for sexual reproduction. During meiosis, meiotic recombination is initiated with programmed DSBs induced by Spo11, and results in the exchange of genetic material between homologous chromosomes, which is beneficial for genetic diversity. Therefore, precise mapping of meiotic DSBs is essential for understanding the mechanism of meiotic homologous recombination. Spo11-induced meiotic DSBs can be divided into three parts: upstream DNA ends, downstream DNA ends, and Spo11-bound oligos.
Both upstream and downstream DNA ends provide 3' overhang structures, which contain a free 3' ssDNA end. There are some disadvantages of previously reported methods for meiotic DSB detection, such as low sensitivity, cumbersome operation, and low resolution. To address these problems, the authors developed the DEtail-seq technique: using Adaptase, a highly efficient single-stranded DNA ligation system, the 3' end of the DNA break is directly ligated to the 1st adaptor, and then the final library is constructed through a series of steps.
After sequencing and data analysis, the position of the 3' end of the DNA break was mapped. The detection of restriction endonuclease cleavage sites showed that the DEtail-seq technique had a very high resolution, at or near the single nucleotide resolution.
Based on the DEtail-seq technique, the investigators profiled the meiotic DSBs in budding yeast, mouse, and human germ cells, and identified novel features of meiotic DSBs. Analysis of the mouse data showed that the meiotic DSB signal at leptotene/zygotene stage was enriched around de novo H3K4me3 peaks at leptotene stage.
Analysis of the human data showed that the meiotic DSB signal was enriched in the CTCF+ enhancer regions. The authors believe that the DEtail-seq technique provides a powerful tool for researching meiosis in various species.
The paper is published in the journal Science China Life Sciences.
More information: Wei Xu et al, DEtail-seq is an ultra-efficient and convenient method for meiotic DNA break profiling in multiple organisms, Science China Life Sciences (2023).
Journal information: Science China Life Sciences
Provided by Science China Press
