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US6096548A Expired / Lapsed US ⏱️ 0.0 years remaining

Method for directing evolution of a virus

Assignee: Codexis Inc
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Filing: 1997-02-03
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Granted: 2000-08-01
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Expires: 2017-02-03
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Abstract

Abstract The invention provides a number of strategies for transferring and/or evolving gene(s) associated with cellular DNA uptake so that they confer or enhance DNA-uptake capacity of a recipient cell. Evolution is achieved by recursive cycles of recombination and screening/selection. One such strategy entails evolving genes that confer competence in one species to confer either greater competence in that species, or comparable or greater competence in a second species. Another strategy entails evolving genes for use as components of cloning vector to confer enhanced uptake of the vector. Other strategies entail evolving viral receptors, viruses, and genes that mediate conjugal transfer.

Inventors

  • 👤 Willem P. C. Stemmer

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-11-04 16:15
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/10; C12N15/1034; C12N15/1058; C12N15/102; C12N15/1027; C12N15/11; C12N15/52; C12N15/63; C12N15/64

Claims Analysis

Total Claims: 16
Claim 1
Independent Claim 179 words
A method of directing evolution of a virus to increase the efficiency with which it infects a host cell, comprising: (1) recombining a DNA segment from the virus to be evolved with at least a second DNA segment, the second DNA segment differing from the first DNA segment in at least two nucleotides, to produce a library of recombinant DNA segments; (2) contacting host cells with a collection of viruses having genomes including the recombinant DNA segments, and screening for viruses that infect the host cells, (3) recombining at least one recombinant DNA segment from a virus infecting the host cells with a further DNA segment, the same or different from the first and second segments, to produce a further library of recombinant DNA segments; (4) contacting host cells with a collection of viruses having genomes including the further recombinant DNA segments, and screening for viruses that infect the cells, (5) repeating (3) and (4), as necessary, until a virus having a genome including a recombinant segment from a further library infects the host cells with a desired efficiency.
Claim 2
Independent Claim 179 words
A method of directing evolution of a virus to increase the efficiency with which it infects a host cell, comprising: (1) recombining a DNA segment from the virus to be evolved with at least a second DNA segment, the second DNA segment differing from the first DNA segment in at least two nucleotides, to produce a library of recombinant DNA segments; (2) contacting host cells with a collection of viruses having genomes including the recombinant DNA segments, and screening for viruses that infect the host cells, (3) recombining at least one recombinant DNA segment from a virus infecting the host cells with a further DNA segment, the same or different from the first and second segments, to produce a further library of recombinant DNA segments; (4) contacting host cells with a collection of viruses having genomes including the further recombinant DNA segments, and screening for viruses that infect the cells, (5) repeating (3) and (4), as necessary, until a virus having a genome including a recombinant segment from a further library infects the host cells with a desired efficiency.
Claim 3
Dependent Claim 18 words
The method of claim 1, wherein the host cell is not naturally susceptible to infection by the virus.
Claim 4
Dependent Claim 29 words
The method of claim 1, wherein the genome of said virus further comprises a selective marker and the viruses are screened by selecting cells that express the selective marker.
Claim 5
Dependent Claim 18 words
The method of claim 1, wherein viruses infecting cells are screened by collecting progeny viruses resulting from infection.
Claim 6
Dependent Claim 22 words
The method of claim 1, wherein the host cells are spread in a layer and the viruses are screened by detecting plaques.
Claim 7
Dependent Claim 22 words
The method of claim 1, further comprising inserting each recombinant DNA segment into a genome of the virus after a recombination step.
Claim 8
Dependent Claim 14 words
The method of claim 2, wherein the DNA segment is a whole viral genome.
Claim 9
Independent Claim 94 words
A method of directing evolution of a virus to increase the efficiency with which it infects a host cell, comprising: conducting a multi-cyclic polynucleotide extension process on overlapping segments having sequences from a population of variants of a polynucleotide encoding at least part of the genome of the virus under conditions whereby one segment serves as a template for extension of another segment to generate a population of recombinant polynucleotides; screening or selecting viruses having a genome comprising one of the recombinant polynucleotides for improved inactivity of the host cell relative to the virus.
Claim 10
Independent Claim 94 words
A method of directing evolution of a virus to increase the efficiency with which it infects a host cell, comprising: conducting a multi-cyclic polynucleotide extension process on overlapping segments having sequences from a population of variants of a polynucleotide encoding at least part of the genome of the virus under conditions whereby one segment serves as a template for extension of another segment to generate a population of recombinant polynucleotides; screening or selecting viruses having a genome comprising one of the recombinant polynucleotides for improved inactivity of the host cell relative to the virus.
Claim 11
Dependent Claim 18 words
The method of claim 8, wherein the host cell is not naturally susceptible to infection by the virus.
Claim 12
Dependent Claim 13 words
The method of claim 8, wherein the polynucleotide encodes a whole viral genome.
Claim 13
Dependent Claim 29 words
The method of claim 8, wherein the genome of the virus further comprises a selective marker and the viruses are screened by selecting cells that express the selective marker.
Claim 14
Dependent Claim 22 words
The method of claim 8, wherein viruses infecting cells are screened by collecting progeny viruses resulting from infection of the host cell.
Claim 15
Dependent Claim 22 words
The method of claim 8, wherein the host cells are spread in a layer and the viruses are screened by detecting plaques.
Claim 16
Dependent Claim 22 words
The method of claim 8, further comprising inserting each recombinant polynucleotide into a genome of the virus before the screening or selecting.

USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 2003-08-01 2004-01-31 2004-08-01 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 2007-08-01 2008-01-31 2008-08-01 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 2011-08-01 2012-01-31 2012-08-01 $7700.00 Window Closed / Lapsed

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