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Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
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Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells
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Ultrasonic Control of Protein Splicing by Split Inteins.

Chuanjiang He1,2, Yu Zhou1,3, Junlin Chen4

  • 1DWI-Leibniz Institute for Interactive Materials, Forckenbeckstr. 50, 52074 Aachen, Germany.

Journal of the American Chemical Society
|September 18, 2024
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Summary
This summary is machine-generated.

Researchers developed an ultrasound-activated protein splicing system for precise control over protein functions. This sonogenetic tool enables remote activation of protein ligation, offering new possibilities for therapeutic applications.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biotechnology

Background:

  • Sonopharmacology and sonogenetics utilize ultrasound to modulate biological processes.
  • Controlling protein activity remotely is crucial for therapeutic interventions.

Purpose of the Study:

  • To develop an ultrasound-responsive protein splicing system for spatiotemporal control of protein ligation.
  • To demonstrate the system's ability to control protein synthesis and cellular signaling.

Main Methods:

  • Engineered split inteins were caged and activated by ultrasound-triggered thrombin release.
  • A DNA-based carrier system was used to deliver the caged components.
  • The system's efficacy was tested using superfolder green fluorescence protein (GFP) and calcitonin synthesis in vitro.

Main Results:

  • Achieved spatiotemporal control of split-intein-mediated protein ligation using focused ultrasound.
  • Successfully synthesized GFP and calcitonin in a controlled manner.
  • Triggered calcitonin receptor-mediated signal transduction in cells without affecting cell viability.

Conclusions:

  • The developed system provides a general method for ultrasound-controlled protein function.
  • This expands the sonogenetic toolbox with protein splicing technologies.
  • Offers a potential pathway for remote control of protein functions in deep tissues.