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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Cell Biology

Background:

  • The discovery of diverse RNA functions and novel RNA molecules is expanding.
  • Current methods for accessing and manipulating endogenous RNAs within live cells are limited.
  • Visualizing specific RNA targets in real-time within living systems remains a challenge.

Purpose of the Study:

  • To develop a universal and sensitive technique for labeling and visualizing endogenous RNAs in live cells.
  • To overcome limitations of existing RNA detection and manipulation methods.
  • To enable the study of natural RNA molecules in their native cellular environment.

Main Methods:

  • Utilized a split aptamer approach combined with fluorescent protein complementation.
  • Designed RNA probes with split aptamer sequences and antisense RNA target sequences.
  • Assembled probes on target RNA to form fluorescent ribonucleoprotein (RNP) complexes for detection.

Main Results:

  • Successfully visualized endogenous bacterial messenger RNAs (mRNAs) in living bacterial cells.
  • Demonstrated highly sensitive RNA detection through the formation of fluorescent RNP complexes.
  • Enabled the detection of chromosomally low-level expressed, unmodified bacterial mRNA.

Conclusions:

  • The developed method provides a universal technique for labeling natural RNAs in live cells.
  • This approach offers sensitive detection and visualization of endogenous RNAs, including low-abundance mRNAs.
  • The technique holds significant potential for basic research, diagnostics, and therapeutics.