Institute of Biotechnology,
CRISPR-Cas nucleases as exemplified by Cas9 and Cas12 are widely used for genome editing applications and are rapidly advancing into the clinics for the treatment of different diseases. However, there are still challenges that need to be overcome. The first challenge is targeting space limitation due to the PAM requirement. I will show how systematic exploration of novel Cas9 orthologues that recognize different PAM sequences allowed us to expand targeting space . Next, I will introduce you miniature nucleases that may help to overcome delivery problems using AAV vectors . And finally, I will talk on the smallest genome editing tools (TnpB) that originates from transposons . Taken together these tools expand genome editing toolbox and provide a versatile platform for different genome manipulations.
- Gasiunas G, et al. A catalogue of biochemically diverse CRISPR-Cas9 orthologs. Nat Commun. 2020 Nov 2;11(1):5512.
- Bigelyte G, et al. Miniature type V-F CRISPR-Cas nucleases enable targeted DNA modification in cells. Nat Commun. 2021;12(1):6191.
- Karvelis T, Druteika G, Bigelyte G, Budre K, Zedaveinyte R, Silanskas A, Kazlauskas D, Venclovas Č, Siksnys V. Transposon-associated TnpB is a programmable RNA-guided DNA endonuclease. Nature. 2021; 599(7886):692-696.