Protospacer integration into plasmid DNA by Cas1–Cas2

We report the integration site specificity of Cas1–Cas2 into target plasmid DNA encoding a CRISPR locus. The goal of this study is to determine the effect of IHF on target site selection of Cas1–Cas2. Methods: After DNA integration reactions, the target plasmid products were fragmented, end-repaired, adapter ligated, PCR amplified and analyzed by Illumina sequencing. Protospacer-containing reads were selected by Cutadapt and mapped to the plasmid with Bowtie

本研究报道了Cas1–Cas2对携带CRISPR位点的靶质粒DNA的整合位点特异性。本研究的目标是明确整合宿主因子IHF（Integration Host Factor）对Cas1–Cas2靶位点选择的影响。实验方法：DNA整合反应完成后，对靶质粒产物依次进行片段化、末端修复、接头连接、PCR扩增，随后通过Illumina测序进行分析。通过Cutadapt筛选出包含原间隔序列（Protospacer）的测序读段，利用Bowtie将其比对至质粒参考序列。