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. Protein/RNA Lego: Pumilio and single‐stranded guide RNAs provide suitable building blocks for programmable self‐assembly of RNP complexes consisting of up to six different components. Correct RNP assembly triggers protein complementation, thereby resulting in increased fluorescence, and is strictly dependent on the correct RNA sequences.