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US 9,023,594 B2Granted
Continuous directed evolution of proteins and nucleic acids
AssigneePresident and Fellows of Harvard College
InventorsDavid R. Liu, Kevin M. Esvelt
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Abstract
The present invention discloses generalizable methods of evolving nucleic acids and proteins utilizing continuous directed evolution. The invention discloses methods of passing a nucleic acid from cell to cell in a desired function-dependent manner. The linkage of the desired function and passage of the nucleic acid from cell to cell allows for continuous selection and mutation of the nucleic acid.
Claims19
A method of continuous evolution of nucleic acids comprising:
- (i) introducing a selection phagemid comprising a gene to be evolved into a flow of bacterial host cells through a lagoon,
- wherein the host cells comprise phage genes required to package the selection phagemid into infectious phage particles,
- wherein at least one gene required to package the selection phagemid into infectious phage particles is expressed in response to expression of the gene to be evolved in the host cell, and wherein the flow rate of the host cells through the lagoon permits replication of the phagemid, but not of the host cells, in the lagoon;
- (ii) replicating and mutating the phagemid within the flow of host cells;
- and
- (iii) isolating a phagemid comprising a mutated gene to be evolved from the flow of cells.
3 more elements
Dependencies are inferred from the claim text.