Pooled CRISPRi screening of the cyanobacterium Synechocystis sp. PCC 6803 for enhanced growth, tolerance, and chemical production

An inducible CRISPRi repression library targeting all annotated genes in the cyanobacterium Synechocystis sp. PCC 6803 was subjected to competitive growth experiments, e.g. under different light regimes and lactate stress, and tracked with high throughput sequencing. Transcriptomics (RNA-Seq) was carried out on certain CRISPRi mutants. Droplet microfluidics screening followed by high throughput sequencing was performed on a lactate-producing iteration of the library. Library amplicon sequencing data has been quality-filtered using Sickle.