Directed Evolution

Directed evolution is a laboratory method for improving genes or the proteins they encode by reproducing the principles of natural selection. Researchers generate genetic diversity through techniques such as mutagenesis, screen or select variants for a desired trait, and repeat these cycles to accumulate beneficial changes. In biology, directed evolution helps optimize enzyme activity, stability, specificity, and other molecular properties that may be difficult to design rationally. The approach supports protein engineering, biotechnology, drug development, and synthetic biology, enabling the creation of catalysts and biological systems with useful functions for research, medicine, and industry.

Directed Evolution - Related Videos

Research

JoVE Journal - Biology
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Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli

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Cited by 12 •

2011

Here we demonstrate a simple protocol to create a random mutant library for a given target sequence. We show how this method, which is performed in vivo in Escherichia coli, can be coupled with functional selections to evolve new enzymatic activities.

Research

JoVE Journal - Biology

Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening

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Cited by 10 •

2016

We present a detailed protocol to construct and screen mutant libraries for directed evolution campaigns in Saccharomyces cerevisiae.

A New Screening Method for the Directed Evolution of Thermostable Bacteriolytic Enzymes

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Cited by 36 •

2012

A novel directed evolution method specific to the field of thermostability engineering was developed and consequently validated for bacteriolytic enzymes. After only one round of random mutagenesis, an evolved bacteriolytic enzyme, PlyC 29C3, displayed greater than twice the residual activity when compared to the wild-type protein after elevated temperature incubation.

Research

JoVE Journal - Genetics
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In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

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2020

Restriction endonucleases with new sequence specificity can be developed from enzymes recognizing a partially degenerate sequence. Here we provide a detailed protocol that we successfully used to alter the sequence specificity of NlaIV enzyme. Key ingredients of the protocol are the in vitro compartmentalization of the transcription/translation reaction and selection of variants with new sequence specificities.

Molecular Evolution of the Tre Recombinase

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Cited by 4 •

2008

Here we report the generation of Tre recombinase through directed, molecular evolution. Tre recombinase recognizes a pre-defined target sequence within the LTR sequences of the HIV-1 provirus, resulting in the excision and eradication of the provirus from infected human cells. While still in its infancy, directed molecular evolution will allow the creation of custom enzymes that will serve as tools of molecular surgery and molecular medicine.

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