CRISPRi as an efficient tool for gene repression in archaea

Aris-Edda Stachler, Thandi Sarah Schwarz, Sandra Schreiber, Anita Marchfelder

Biology II, Ulm University, 89069 Ulm, Germany

Abstract

In the years following its discovery and characterization, the CRISPR-Cas system has been modified and converted into a multitude of applications for eukaryotes and bacteria, such as genome editing and gene regulation. Since no such method has been available for archaea, we developed a tool for gene repression in the haloarchaeon Haloferax volcanii by repurposing its endogenous type I-B CRISPR-Cas system. Here, we present the two possible approaches for gene repression as well as our workflow to achieve and assess gene knockdown, offer recommendations on protospacer selection and give some examples of genes we have successfully silenced.