In vitro evolution in microfluidic systems
View Patent ↗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; 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.
1. A method for detecting a selectable change within a microcapsule, comprising the steps of:
forming a plurality of aqueous microcapsules in a fluorinated oil comprising a fluorinated polymer surfactant, wherein each aqueous microcapsule comprises a cell and a first genetic element;
pooling the aqueous microcapsules such that a portion of the aqueous microcapsules contact each other without fusing together;
incubating the pooled microcapsules to cause the first genetic element to interact with a molecule associated with the cell; and
detecting a selectable change in one or more of the pooled microcapsules.
2. The method of claim 1 , further comprising pooling the microcapsules into one or more common compartments such that a portion of the plurality of microcapsules contact each other but do not fuse.
3. The method of claim 1 , wherein the selectable change comprises an optical signal.
4. The method of claim 1 , wherein the first genetic element comprises one or more primer sequences for amplification.
5. The method of claim 1 , wherein the cell is an immune cell.
6. The method of claim 5 , wherein the immune cell is a B-cell.
7. The method of claim 1 , wherein the molecule is a reporter protein.
8. The method of claim 1 , wherein the molecule comprises a gene.
9. The method of claim 8 , further comprising expressing the gene to form a gene product.
10. The method of claim 9 , wherein the gene product is a protein.
11. The method of claim 10 , wherein the protein is an antibody.
12. The method of claim 10 , wherein the protein is located within the cell.
13. The method of claim 10 , wherein the protein is located outside of the cell.
14. The method of claim 9 , wherein the gene product is cDNA.
15. The method of claim 1 , wherein the molecule comprises a gene product having enzymatic or binding activity.
16. The method of claim 1 , wherein the incubation step comprises performing an amplification reaction.
17. The method of claim 16 , wherein the amplification reaction is a polymerase chain reaction (PCR).
18. The method of claim 1 , wherein the molecule is DNA or RNA.
19. The method of claim 1 , wherein the detecting step comprises performing a sequencing reaction.