CRISPRi screen in E. coli MG1655 with the EcoWG1 library. EcoWG1

Gene silencing with dCas9 is quickly becoming a standard method to study and engineer bacteria1–4, yet the design rules that determine on-target activity are still unknown. Taking advantage of high-throughput screening data, we fit a model able to predict dCas9’s ability to block the RNA polymerase. Using this model we further build a novel genome-wide library for E. coli MG1655, EcoWG1, and demonstrate its improved performance over published datasets.

利用失活Cas9（dCas9）进行基因沉默正迅速成为研究与工程改造细菌的标准方法1–4，但决定其靶点活性的设计规则仍未明确。本研究借助高通量筛选数据，拟合得到可预测dCas9阻断RNA聚合酶能力的模型。基于该模型，我们进一步为大肠杆菌MG1655构建了全新的全基因组文库EcoWG1，并证实其性能优于已发表的相关数据集。