Optical sorting method
First Claim
1. A method for isolating one or more genetic elements encoding a gene product having a desired activity the expression of which may result, directly or indirectly, in the modification of an optical property of a genetic element encoding the gene product, comprising the steps of:
- (a) expressing the genetic elements to produce their respective gene products such that the gene products are linked to the genes encoding them;
(b) compartmentalising genetic elements into microcapsules; and
(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.
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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.
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Citations
44 Claims
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1. A method for isolating one or more genetic elements encoding a gene product having a desired activity the expression of which may result, directly or indirectly, in the modification of an optical property of a genetic element encoding the gene product, comprising the steps of:
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(a) expressing the genetic elements to produce their respective gene products such that the gene products are linked to the genes encoding them;
(b) compartmentalising genetic elements into microcapsules; and
(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. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41)
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42. A method for preparing a gene product, comprising the steps of:
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(a) preparing a genetic element encoding the gene product;
(b) expressing the genetic elements to produce their respective gene products;
(c) compartmentalising genetic elements into 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.
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43. A method for screening a compound or compounds capable of modulating the activity of a gene product, comprising the steps of:
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(a) preparing a genetic element encoding the gene product;
(b) expressing the genetic elements to produce their respective gene products;
(c) compartmentalising genetic elements into 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.
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44. A method for preparing a compound or compounds comprising the steps of:
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(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 this step;
(c) expressing the genetic elements to produce their respective gene products;
(d) compartmentalising the genetic elements into 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 the relevant step of the synthesis.
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Specification