Engineered, nucleocytoplasmic shuttling Cas13d enables highly efficient cytosolic RNA targeting
Christoph Gruber, Lea Krautner, Valter Bergant, Vincent Grass, Zhe Ma, Lara Rheinemann, Ariane Krus, Friederike Reinhardt, Lyupka Mazneykova, Marianne Rocha-Hasler, Dong‐Jiunn Jeffery Truong, Gil G. Westmeyer, Andreas Pichlmair, Gregor Ebert, Florian Giesert, Wolfgang Wurst
Abstract
CRISPR/Cas13 systems are programmable tools for manipulating RNAs and are used in a variety of RNA-targeting applications 1 , 2 , 3 . Within the Cas13 family, Cas13d is the most active subtype in mammalian cells 4 , 5 . Recently, Cas13d was harnessed as an antiviral against diverse human RNA viruses 6 , 7 . However, Cas13d is barely active in the cytosol of mammalian cells, restricting its activity to the nucleus, which limits applications such as programmable antivirals 4 , 5 . Most RNA viruses replicate exclusively in the cytosol, suggesting that current Cas13d-based antivirals rely on uncontrolled nuclear leakage and are therefore limited in their efficiency 7 .