Optical sorting method
AssigneeMedical Research Council
InventorsAndrew Griffiths, Dan Tawfik, Armin Sepp
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Abstract
The invention describes a method for isolating one or more genetic elements encoding a gene product having a desired activity, comprising the steps of: (a) compartmentalising genetic elements into microcapsules; (b) expressing the genetic elements to produce their respective gene products within the microcapsules; (c) sorting the genetic elements which produce the gene product having the desired activity using a change in the optical properties of the genetic elements. The invention enables the in vitro evolution of nucleic acids and proteins by repeated mutagenesis and iterative applications of the method of the invention.
Claims43
A method for isolating one or more genetic elements encoding a gene product having a desired activity the expression of which results, directly or indirectly, in the modification of an optical property of a genetic element encoding the gene product, comprising the steps of:
- (a) compartmentalising genetic elements into microcapsules;
- (b) expressing the genetic elements to produce their respective gene products within the microcapsules;
- (c) sorting the genetic elements which produce the gene product(s) having the desired activity according to a change in the optical properties of the genetic elements, whereby a genetic element encoding a gene product having said desired activity is isolated.
A method for preparing a gene product, comprising the steps of:
- (a) preparing plurality of a genetic elements encoding a plurality of gene products;
- (b) compartmentalising said genetic elements into microcapsules;
- (c) expressing the genetic elements to produce their respective gene products within the microcapsules;
- (d) sorting the genetic elements which produce the gene product(s) having the desired activity using a change in their optical properties;
- and
- (e) expressing the gene product having the desired activity.
A method for screening a compound or compounds capable of modulating the activity of a gene product, comprising the steps of:
- (a) preparing a repertoire of genetic elements encoding gene products;
- (b) compartmentalising the genetic elements into microcapsules;
- (c) expressing the genetic elements to produce their respective gene products within the microcapsules;
- (d) sorting the genetic elements which produce the gene product(s) having the desired activity using a change in their optical properties;
- and
- (e) contacting a gene product having the desired activity with the compound or compounds and monitoring the modulation of an activity of the gene product by the compound or compounds.
In a method of preparing a compound, the method comprising a step of contacting a polypeptide with a reagent, wherein said contacting facilitates at least one step in said method of preparing a compound, the improvement comprising:
- a) preparing genetic elements encoding variants of said polypeptide;
- b) compartmentalizing said genetic elements into microcapsules;
- c) expressing the genetic elements to produce their respective gene products within the microcapsules;
- d) sorting a genetic element which produces a polypeptide gene product having a desired activity using a change in an optical property of the genetic element to sort, wherein a gene product having said desired activity is identified;
- and
- e) using the polypeptide identified in step (d) in said method of preparing a compound.
Dependencies are inferred from the claim text.