Robust single-cell discovery of RNA targets of RNA-binding proteins and ribosomes

Kristopher W Brannan1,2,3, Isaac A Chaim1,2,3, Ryan J Marina1,2,3, Brian A Yee1,2,3, Eric R Kofman1,2,3, Daniel A Lorenz1,2,3, Pratibha Jagannatha1,2,3, Kevin D Dong1,2,3, Assael A Madrigal1,2,3, Jason G Underwood4, Gene W Yeo5,6,7

  1. Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, CA, USA.
  2. Stem Cell Program, University of California San Diego, La Jolla, CA, USA.
  3. Institute for Genomic Medicine, University of California San Diego, La Jolla, CA, USA.
  4. Pacific Biosciences (PacBio) of California, Menlo Park, CA, USA.
  5. Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, CA, USA. geneyeo@ucsd.edu.
  6. Stem Cell Program, University of California San Diego, La Jolla, CA, USA. geneyeo@ucsd.edu.
  7. Institute for Genomic Medicine, University of California San Diego, La Jolla, CA, USA. geneyeo@ucsd.edu.

Abstract

RNA-binding proteins (RBPs) are critical regulators of gene expression and RNA processing that are required for gene function. Yet the dynamics of RBP regulation in single cells is unknown. To address this gap in understanding, we developed STAMP (Surveying Targets by APOBEC-Mediated Profiling), which efficiently detects RBP-RNA interactions. STAMP does not rely on ultraviolet cross-linking or immunoprecipitation and, when coupled with single-cell capture, can identify RBP-specific and cell-type-specific RNA-protein interactions for multiple RBPs and cell types in single, pooled experiments. Pairing STAMP with long-read sequencing yields RBP target sites in an isoform-specific manner. Finally, Ribo-STAMP leverages small ribosomal subunits to measure transcriptome-wide ribosome association in single cells. STAMP enables the study of RBP-RNA interactomes and translational landscapes with unprecedented cellular resolution.

Presented By Kristopher W Brannan | ORCID iD

Biochemistry, Biotechnology, Genetics & Genome Biology, GenomicsNicholas Liao2021-05 (May)