Team:NTU-Singapore/Basic Part

Basic Parts

Truncated dCas9

BBa_K2130000 ∆RuvCIII-2 ∆HNH SP-dCas9

This tuncated version of dCas9 is the smallest dCas9 that works the best. The ∆RuvCIII-2 truncation has been rationally predicted by comparison of crystal structures between two states of Cas9.

Conformations of RuvCIII in state A, before NTS cleavage and state B, after NTS cleavage

Schematic of RuvCIII sub-domain deletions

It is able to retain ~70% of binding strength with 15% of the protein is deleted.

Activation of a zsGreen reporter gene using the various RuvCIII truncations

Microscope image of HEK293FT cells 24 hours after transfection.