Bennett-Baker Pamela E, Mueller Jacob L
Department of Human Genetics, University of Michigan Medical School, Ann Arbor, MI 48109, USA.
Nucleic Acids Res. 2017 Nov 2;45(19):e165. doi: 10.1093/nar/gkx749.
Megabase-sized, complex, repetitive regions of genomes are poorly studied, due to the technical and computational challenges inherent to both assembling precise reference sequences and accurately assessing structural variation across contiguous megabase DNA regions. Here we describe a strategy to overcome these challenges, CISMR (CRISPR-mediated isolation of specific megabase-sized regions of the genome), which enables us to perform targeted isolation of contiguous megabase-sized segments of the genome. Direct sequencing of the purified DNA segments can have >100-fold enrichment of the target region, thus enabling the exploration of both DNA sequence and structural diversity of complex genomic regions in any species.
由于在组装精确的参考序列以及准确评估跨越连续兆碱基DNA区域的结构变异方面存在固有的技术和计算挑战，基因组中兆碱基大小的复杂重复区域研究较少。在此，我们描述了一种克服这些挑战的策略——CISMR（CRISPR介导的基因组特定兆碱基大小区域分离），它使我们能够对基因组中连续的兆碱基大小片段进行靶向分离。对纯化的DNA片段进行直接测序可使目标区域富集100倍以上，从而能够探索任何物种复杂基因组区域的DNA序列和结构多样性。
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