Deaminase-based RNA recording enables high throughput mutational profiling of protein-RNA interactions

Engineered TadA8.20 deaminase enables RNA recording of protein-RNA recognition, recovering key boxB nucleotides and λN residues via nucleotide edits read by DNA sequencing.

• Why it matters: Protein-RNA interactions are central to RNA metabolism and often dysregulated in disease, but scalable ways to link RNA and protein sequence requirements across settings are limited.
• What they did: Using the λN-boxB complex, the study performs pooled mutational scanning on RNA and protein, then compares boxB libraries in vitro versus in human cells, including direct fusion and split recruitment via nanobody and protein A/G fusions.
• The result: Sequence requirements for protein-RNA recognition are conserved across molecular and cellular contexts, and fusion or split deaminase recruitment preserves similar patterns of sequence discrimination.

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