Genetically Encoded Sensor Enables Endogenous RNA Imaging with Conformation-Switching Induced Fluorogenic Proteins

J Am Chem Soc. 2021 Sep 8;143(35):14394-14401. doi: 10.1021/jacs.1c07719. Epub 2021 Aug 25.

Abstract

Genetically encoded molecular tools are crucial for live cell RNA imaging, and few are available for endogenous RNA imaging. We develop a new genetically encoded sensor using conformation switching RNA induced fluorogenic proteins that enable multicolor and signal-amplified imaging of endogenous RNAs. The sensor system is designed with an RNA sensing module and a degron-fused fluorescent protein reporter. Target RNA induces conformation switching of the RNA sensing module to form RNA aptamers that stabilize the degron-fused protein for fluorogenic imaging. This sensor is demonstrated for high-contrast imaging of survivin mRNA abundance and dynamics in live cells. Moreover, the sensor system is extended to a multicolor palette by screening fluorogenic proteins of distinct colors, and engineered into a signal amplifier using the split fluorescent protein design. The sensor is further exploited for imaging lncRNA MALAT-1 and its translocation dynamics during mitosis. Our sensor system can afford a valuable platform for RNA imaging in biomedical research and clinical theranostics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aptamers, Nucleotide / analysis*
  • Aptamers, Nucleotide / chemistry
  • Aptamers, Nucleotide / genetics
  • Cell Line, Tumor
  • Green Fluorescent Proteins / chemistry*
  • Humans
  • Nucleic Acid Hybridization
  • Optical Imaging / methods
  • Protein Conformation
  • RNA, Long Noncoding / analysis*
  • RNA, Long Noncoding / chemistry
  • RNA, Long Noncoding / genetics
  • RNA, Messenger / analysis*
  • RNA, Messenger / chemistry
  • RNA, Messenger / genetics
  • Survivin / genetics

Substances

  • Aptamers, Nucleotide
  • RNA, Long Noncoding
  • RNA, Messenger
  • Survivin
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins