We firslt apply
protein structure analysis to find the best activation domain for CRISPRa, the results showed that
the α domain of RNAp from Vibrio natriegens could accomplish the best gene
activation compared with others.
So we firstly fusion
V.nαdomain in the N domain of dCasΦ，but it impeded the gene expression while classic AD SoxS showed
a successful activation. We assumed that this may due to the fusion manner which was then proved by
both docking analysis and practical function tests that Cas-linker-AD is better
than our first design, AD-linker-Cas.
comparing the modeling results from the I-TASSER server, we selected a relatively more
appropriate linker from the six linkers collected.
Fig 1. Integration of the protein structure model and our design.