In vitro evolution in microfluidic systems
First Claim
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1. A method for producing an encapsulated gene product, comprising the steps of:
- (a) providing within a first microcapsule within a microfluidic channel a first genetic element encoding a first gene product, the microcapsule comprising an aqueous fluid substantially surrounded by an immiscible fluorinated oil including a fluorinated polymer surfactant, wherein the microcapsule is able to contact without fusing with another microcapsule due to presence of the surfactant;
(b) providing conditions within the first microcapsule suitable for the expression of the first genetic element to form its first gene product;
(c) introducing at least one of a first microbead to the first microcapsule, wherein the first microbead is either encapsulated within the first microcapsule prior to the expression of the first genetic element or is encapsulated within the first microcapsule subsequent to the expression of the first genetic element; and
(d) complexing the first gene product with at least one of the plurality of first microbeads via a capture moiety on the first microbead.
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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) compartmentalizing genetic elements into microcapsules; and (b) sorting the genetic elements which express the gene product having the desired activity; wherein at least one step is under microfluidic control. 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
38 Claims
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1. A method for producing an encapsulated gene product, comprising the steps of:
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(a) providing within a first microcapsule within a microfluidic channel a first genetic element encoding a first gene product, the microcapsule comprising an aqueous fluid substantially surrounded by an immiscible fluorinated oil including a fluorinated polymer surfactant, wherein the microcapsule is able to contact without fusing with another microcapsule due to presence of the surfactant; (b) providing conditions within the first microcapsule suitable for the expression of the first genetic element to form its first gene product; (c) introducing at least one of a first microbead to the first microcapsule, wherein the first microbead is either encapsulated within the first microcapsule prior to the expression of the first genetic element or is encapsulated within the first microcapsule subsequent to the expression of the first genetic element; and (d) complexing the first gene product with at least one of the plurality of first microbeads via a capture moiety on the first microbead. - 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)
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Specification