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In vitro Mutagenesis01:16

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To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
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Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
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A Protocol for Functional Assessment of Whole-Protein Saturation Mutagenesis Libraries Utilizing High-Throughput Sequencing
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The Parasol Protocol for computational mutagenesis.

P G A Aronica1, C Verma2, B Popovic3

  • 1Department of Chemistry and Institute of Chemical Biology, Imperial College London, South Kensington Campus, Exhibition Road, London SW7 2AZ, UK.

Protein Engineering, Design & Selection : PEDS
|June 4, 2016
PubMed
Summary
This summary is machine-generated.

We developed the Parasol Protocol for computational site-directed mutagenesis to accelerate therapeutic peptide and protein discovery. This automated method shows good agreement with experimental data for stapled peptides.

Keywords:
in silico mutationmolecular dynamics

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Area of Science:

  • Biochemistry and Molecular Biology
  • Computational Chemistry
  • Drug Discovery

Background:

  • Peptides and proteins are crucial therapeutic agents.
  • Predictive modeling aids in their discovery and development.
  • Computational methods can optimize research workflows.

Purpose of the Study:

  • To introduce the Parasol Protocol, a novel computational tool.
  • To enable efficient, automated site-directed mutagenesis.
  • To validate the protocol's accuracy in predicting experimental outcomes.

Main Methods:

  • Developed the Parasol Protocol using the AMBER Molecular Dynamics (MD) package.
  • Implemented a dynamic method for computational site-directed mutagenesis.
  • Automated the mutation process between any amino acid pairs.

Main Results:

  • The Parasol Protocol facilitates computationally expedient mutagenesis.
  • Demonstrated the protocol's utility with a stapled peptide case study.
  • Achieved good agreement between computational predictions and experimental results.

Conclusions:

  • The Parasol Protocol is a valuable tool for therapeutic peptide and protein research.
  • This method can significantly direct and accelerate drug discovery workflows.
  • Computational mutagenesis offers a powerful approach to experimental validation.