Modular assembly of designer PUF proteins for specific post-transcriptional regulation of endogenous RNA.

ABSTRACT:

Background

Due to their modular repeat structure, Pumilio/fem-3 mRNA binding factor (PUF) proteins are promising candidates for designer RNA-binding protein (RBP) engineering. To further facilitate the application of the PUF domain for the sequence-specific RBP engineering, a rapid cloning approach is desirable that would allow efficient introduction of multiple key amino acid mutations in the protein. Here, we report the implementation of the Golden Gate cloning method for an efficient one-step assembly of a designer PUF domain for RNA specificity engineering.

Results

We created a repeat module library that is potentially capable of generating a PUF domain with any desired specificity. PUF domains with multiple repeat modifications for the recognition of altered RNA targets

SUBMITTER: Abil Z 

PROVIDER: S-EPMC3943411 | biostudies-literature | 2014 Mar

REPOSITORIES: biostudies-literature