Genome-wide profiling of histone modifications in Plasmodium falciparum using CUT&RUN.

Genome-wide profiling of histone modifications in Plasmodium falciparum using CUT&RUN.
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DOI:
10.26508/lsa.202201778
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发表时间:
2023-01
影响因子:
4.4
通讯作者:
--
中科院分区:
生物学2区
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我们提出了一种详细的循序渐进的方案,用于使用Cut&Run方法对组蛋白修饰进行可靠的全基因组分析,该方法只需要很少的恶性疟原虫血液阶段。我们最近采用了一种切割-运行方案,用于人类疟疾寄生虫疟原虫染色质修改的全基因组图谱。使用下面描述的循序渐进的方案,我们能够仅使用芯片序列所需的一小部分细胞来生成多个组蛋白修饰的高质量图谱。使用针对两种常见的组蛋白修饰,H3K4me3和H3K9me3的抗体,我们在这里表明,切割和运行图谱具有高度的重复性,并紧密概括了之前发表的基于芯片序列的组蛋白标记丰度图谱。最后,我们表明,与芯片序列相比,切割和运行所需的测序覆盖率要低得多,以实现准确的测序。
We present a detailed step-wise protocol for reliable genome-wide profiling of histone modifications using CUT&RUN that requires only low numbers of Plasmodium falciparum blood stages. We recently adapted a CUT&RUN protocol for genome-wide profiling of chromatin modifications in the human malaria parasite Plasmodium. Using the step-by-step protocol described below, we were able to generate high-quality profiles of multiple histone modifications using only a small fraction of the cells required for ChIP-seq. Using antibodies against two commonly profiled histone modifications, H3K4me3 and H3K9me3, we show here that CUT&RUN profiling is highly reproducible and closely recapitulates previously published ChIP-seq-based abundance profiles of histone marks. Finally, we show that CUT&RUN requires substantially lower sequencing coverage for accurate profiling compared with ChIP-seq.
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