Computational identification of clonal cells in single-cell CRISPR screens.

Computational identification of clonal cells in single-cell CRISPR screens.
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DOI:
10.1186/s12864-022-08359-1
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发表时间:
2022-02-15
期刊:
影响因子:
4.4
通讯作者:
Hon GC
Hon GC
中科院分区:
生物学2区
文献类型:
--
作者:
Wang Y;Xie S;Armendariz D;Hon GC

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单细胞CRISPR筛选是通过将遗传扰动与全转录组表型联系起来来了解基因组功能的强大工具。然而,由于在这些筛选中几乎没有细胞可以负担得起测序,因此细胞的偏倚采样可能影响数据解释。有偏采样的一个潜在来源是克隆细胞扩增。在这里,我们使用多重sgRNA作为条形码在单细胞筛选中鉴定克隆细胞。我们发现每个克隆中的细胞具有转录相似性,并具有片段拷贝数变化。这些分析表明克隆在遗传上是不同的。最后,我们发现克隆扩增细胞的转录相似性导致了单细胞CRISPR筛选中的假阳性。减少克隆细胞的克隆扩增或计算过滤的实验条件将提高单细胞CRISPR筛选的可靠性。在线版本包含补充材料,可通过10.1186/s12864-022-08359-1获得。
Single-cell CRISPR screens are powerful tools to understand genome function by linking genetic perturbations to transcriptome-wide phenotypes. However, since few cells can be affordably sequenced in these screens, biased sampling of cells could affect data interpretation. One potential source of biased sampling is clonal cell expansion. Here, we identify clonal cells in single cell screens using multiplexed sgRNAs as barcodes. We find that the cells in each clone share transcriptional similarities and bear segmental copy number changes. These analyses suggest that clones are genetically distinct. Finally, we show that the transcriptional similarities of clonally expanded cells contribute to false positives in single-cell CRISPR screens. Experimental conditions that reduce clonal expansion or computational filtering of clonal cells will improve the reliability of single-cell CRISPR screens. The online version contains supplementary material available at 10.1186/s12864-022-08359-1.
DOI: 10.1038/nature24284
发表时间: 2017-11-02
期刊: Nature
影响因子: 64.8
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发表时间: 2016-12-15
期刊: CELL
影响因子: 64.5
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