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Pooled CRISPRi screening of the cyanobacterium Synechocystis sp PCC 6803 for enhanced industrial phenotypes.

Abstract：

:Cyanobacteria are model organisms for photosynthesis and are attractive for biotechnology applications. To aid investigation of genotype-phenotype relationships in cyanobacteria, we develop an inducible CRISPRi gene repression library in Synechocystis sp. PCC 6803, where we aim to target all genes for repression. We track the growth of all library members in multiple conditions and estimate gene fitness. The library reveals several clones with increased growth rates, and these have a common upregulation of genes related to cyclic electron flow. We challenge the library with 0.1 M L-lactate and find that repression of peroxiredoxin bcp2 increases growth rate by 49%. Transforming the library into an L-lactate-secreting Synechocystis strain and sorting top lactate producers enriches clones with sgRNAs targeting nutrient assimilation, central carbon metabolism, and cyclic electron flow. In many examples, productivity can be enhanced by repression of essential genes, which are difficult to access by transposon insertion.

journal_name

Nat Commun

journal_title

Nature communications

authors

Yao L,Shabestary K,Björk SM,Asplund-Samuelsson J,Joensson HN,Jahn M,Hudson EP

doi

10.1038/s41467-020-15491-7

subject

Has Abstract

pub_date

2020-04-03 00:00:00

pages

1666

issue

1

issn

2041-1723

pii

10.1038/s41467-020-15491-7

journal_volume

11

pub_type

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