Escherichia coli Raw sequence reads. Escherichia coli

Genetic interaction mapping is useful for understanding the molecular basis of cellular decision making, but elucidating interactions genome-wide is challenging due to the massive number of gene combinations that must be tested. Here, we demonstrate a simple approach to thoroughly map genetic interactions in bacteria using microfluidic-based single cell sequencing. Using single cell PCR in droplets, we link distinct genetic information into single DNA sequences that can be decoded by next generation sequencing. Our approach is scalable and theoretically enables the pooling of entire interaction libraries to interrogate every possible pairwise genetic interaction in a single culture. The speed, ease, and low-cost of our approach makes genetic interaction mapping viable for routine characterization, allowing the interaction network to be used as a universal read out for a variety of biology experiments, and for the elucidation of interaction networks in non-model organisms.

遗传相互作用图谱（genetic interaction mapping）对于解析细胞决策的分子基础具有重要价值，但由于需检测的基因组合数量极为庞大，在全基因组范围内阐明遗传相互作用仍极具挑战性。在此研究中，我们提出了一种基于微流控单细胞测序（microfluidic-based single cell sequencing）的简便方法，可全面绘制细菌的遗传相互作用图谱。通过液滴式单细胞PCR（single cell PCR）技术，我们能够将各异的遗传信息整合为可通过下一代测序（next generation sequencing）解码的单一段DNA序列。该方法具备可扩展性，理论上可将全部相互作用文库进行混合，从而在单一培养体系中检测所有可能的成对遗传相互作用。本方法兼具快速、简便与低成本的优势，使得遗传相互作用图谱可用于常规的样本特征鉴定，进而使相互作用网络可作为多种生物学实验的通用检测读出信号，同时也可用于阐释非模式生物（non-model organisms）的相互作用网络。