Optical sorting method
AssigneeMedical Research Council
InventorAndrew Griffiths
Life of this patent
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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.
Claims44
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.
The method according to claim I further comprising the additional step, after step (c), of:
- (d) introducing one or more mutations into the genetic element(s) isolated in step (c).
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 product;
- (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.
A method for preparing a compound or compounds comprising the steps of:
- (a) providing a synthesis protocol wherein at least one step is facilitated by a polypeptide;
- (b) preparing genetic elements encoding variants of the polypeptide which facilitates said at least one step;
- (c) compartmentalising the genetic elements into microcapsules;
- (d) expressing the genetic elements to produce their respective gene products within the microcapsules;
- (e) sorting the genetic elements which produce polypeptide gene product(s) having the desired activity using a change in their optical properties;
- and
- (f) preparing the compound or compounds using the polypeptide gene product identified in (e) to facilitate said at least one step of the synthesis.
Dependencies are inferred from the claim text.