Vector-based mutagenesis system
AssigneePresident and Fellows of Harvard College
InventorsAhmed Hussein Badran, David R. Liu
Life of this patent
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Abstract
Strategies, reagents, methods, and systems for modulating the mutation rate in cells are provided herein. The strategies, reagents, methods, and systems are broadly applicable for the modulation of mutation rates in cells where high mutation rates and/or control over a broad range of mutation rates is desired, for example, in the context of diversifying a nucleic acid sequence or a plurality of such sequences within a population of cells, for the generation of diversified nucleic acid libraries, and for directed evolution of nucleic acids and encoded products.
Claims26
An expression construct for modulating the mutation rate of nucleic acids in a bacterial cell, the construct comprising a nucleic acid sequence encoding:
- (a) at least one gene product that disrupts a proofreading pathway, a translesion synthesis pathway, and/or a methyl-directed mismatch repair pathway of the bacterial cell;
- and
- (b) at least one gene product that disrupts a base excision repair pathway and/or a base selection pathway of the bacterial cell;
- wherein the gene products are under the control of one or more heterologous promoters.
An expression construct comprising a nucleic acid sequence under the control of a heterologous promoter, wherein the expression construct comprises the sequence set forth in any one of SEQ ID NO:
- 27, 28, 29, 33, 34, and 35.
Dependencies are inferred from the claim text.