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Area of Science:

  • Molecular Biology
  • RNA Biology
  • Biochemistry

Background:

  • Dicer and its partners TRBP and PACT are crucial for regulatory RNA biogenesis.
  • One strand of diced dsRNA is preferentially loaded into RISC.
  • The mechanism of strand selection and Dicer's role remain unclear.

Purpose of the Study:

  • To elucidate the mechanism of strand selection during RISC loading in humans.
  • To investigate the role of Dicer in sensing dsRNA thermodynamic properties.
  • To understand how Dicer interacts with dsRNA and its partners.

Main Methods:

  • Analysis of dsRNA repositioning within Dicer complexes post-dicing.
  • Investigating Dicer's ability to sense siRNA thermodynamic asymmetry.
  • Studying Dicer-dsRNA interactions with and without dsRBPs.

Main Results:

  • dsRNAs significantly reposition within Dicer after dicing.
  • This repositioning facilitates directional binding and strand selection based on helix thermodynamics.
  • Dicer can sense siRNA thermodynamic asymmetry independently of TRBP or PACT.

Conclusions:

  • Dicer plays an active role in guide strand selection by sensing thermodynamic asymmetry.
  • Dicer utilizes distinct RNA-binding sites for processing and thermodynamic sensing.
  • A model is proposed where Dicer directs RISC loading through dual-site binding during siRNA biogenesis.