CRISPRi-based functional genomic screening identifies genes essential for CH 4 -dependent growth in a methanotrophic bacterium

Methanotrophic bacteria are the primary organisms that consume atmospheric methane (CH 4 ) and have potential to mitigate climate-active gases. However, a limited understanding of the genetic determinants of methanotrophy hinders the development of biotechnologies leveraging these unique microbes. Here, we developed and optimized a CRISPR interference (CRISPRi) system to enable functional genomic screening in methanotrophic bacteria. We built a genome-wide single guide RNA (sgRNA) library in the