Programmable Design of Functional Ribonucleoprotein Complexes

Ribonucleoprotein (RNP) complexes are widespread in nature and play crucial roles in gene regulation, RNA processing, and translation. Novel technologies, such as CRISPR-mediated genome engineering, stress the potential of RNP complexes to carry out complex tasks in molecular biology. Here we report a bottom-up approach for the programmable self-assembly of RNP complexes. The building blocks for RNP complex formation are RNAs and Pumilio proteins that can bind to RNA sequence-specifically. Correct RNP assembly triggers protein complementation of a tripartite GFP, thereby resulting in up to 25-fold increased fluorescence, and is strictly dependent on the correct RNA sequences. Our results indicate that Pumilio and guide RNAs are suitable building blocks for the correct self-assembly of RNP complexes consisting of up to six different components. Self-assembling RNP complexes might prove useful for complex biotechnological applications in RNA sensing, imaging, or processing.