Massively parallel high-order combinatorial genetics in human cells.
Massively parallel high-order combinatorial genetics in human cells.
复制标题
DOI：
10.1038/nbt.3326
复制
发表时间：
2015-09
影响因子：
46.9
通讯作者：
Lu TK
中科院分区：
文献类型：
--
作者：
Wong AS;Choi GC;Cheng AA;Purcell O;Lu TK
组合遗传学的系统功能分析一直受限于所能达到的通量和所能研究的复杂性的顺序。为了实现人类细胞中基因组合的大规模并行表征，我们开发了一种快速、可扩展组装高阶条形码组合遗传文库的技术，该技术可以通过高通量测序进行量化。我们应用这种组合遗传学技术（CombiGEM），创建了39个人类microRNA （miRNA）前体的1,521个双导和51,770个三导条形码组合的高覆盖率文库。我们确定了miRNA组合，协同使耐药癌细胞对化疗敏感和/或抑制癌细胞增殖，为复杂的miRNA网络提供了见解。更广泛地说，我们的方法将使调节与生物医学、生物技术和基础科学相关的表型的多因子遗传组合的高通量分析成为可能。
The systematic functional analysis of combinatorial genetics has been limited by the throughput that can be achieved and the order of complexity that can be studied. To enable massively parallel characterization of genetic combinations in human cells, we developed a technology for rapid, scalable assembly of high-order barcoded combinatorial genetic libraries that can be quantified with high-throughput sequencing. We applied this technology, combinatorial genetics en masse (CombiGEM), to create high-coverage libraries of 1,521 two-wise and 51,770 three-wise barcoded combinations of 39 human microRNA (miRNA) precursors. We identified miRNA combinations that synergistically sensitize drug-resistant cancer cells to chemotherapy and/or inhibit cancer cell proliferation, providing insights into complex miRNA networks. More broadly, our method will enable high-throughput profiling of multifactorial genetic combinations that regulate phenotypes of relevance to biomedicine, biotechnology and basic science.