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Chapters
- 00:05Introduction
- 00:45What is your conceptual approach?
- 01:29Mechanisms of action
- 02:38Identifying a target site
- 03:04Principles of molecular evolution
- 04:27Mutation and selection
- 06:06Defining a “new” recombinase?
- 06:45Dynamics of the evolutionary process
- 08:11Potential limitations
- 08:54Functional stability of the Tre recombinase
- 09:23Steps towards therapeutic use
- 10:27Molecular medicine
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.
Tags
Molecular EvolutionTre RecombinaseDirected EvolutionHIV-1 ProvirusCre RecombinaseLoxP SequenceLTR SequencesGenomic SequencesRecombination ActivityMutagenized Cre LibrariesDirected Molecular Evolution ProcessResidual ActivityAsymmetric LoxLTR SequencesRecombinase LibrariesReiterative Evolution CyclesShuffled And Recombined LibrariesSynergistic Mutations
