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US 12,398,390 B2Granted
Negative selection and stringency modulation in continuous evolution systems
AssigneePresident and Fellows of Harvard College
InventorsDavid R. Liu, Jacob Charles Carlson, Ahmed Hussein Badran, Kevin Michael Esvelt
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Abstract
Strategies, systems, methods, reagents, and kits for phage-assisted continuous evolution are provided herein. These include strategies, systems, methods, reagents, and kits allowing for stringency modulation to evolve weakly active or inactive biomolecule variants, negative selection of undesired properties, and/or positive selection of desired properties.
Claims20
A vector system comprising:
- a selection phagemid, a low selection stringency plasmid, and a high selection stringency plasmid, wherein
- the low selection stringency plasmid comprises a first nucleic acid encoding a first pIII protein that is operably linked to a small molecule inducible promoter;
- the high selection stringency plasmid comprises a second nucleic acid encoding a second pIII protein that is operably linked to an activity-dependent promoter comprising a binding site for a DNA-binding gene product;
- and
- the selection phagemid comprises a third nucleic acid encoding the DNA-binding gene product.
2 more elements
Dependencies are inferred from the claim text.