Evolution of whole cells and organisms by recursive sequence recombination
View Patent ↗The invention provides methods employing iterative cycles of recombination and selection/screening for evolution of whole cells and organisms toward acquisition of desired properties Examples of such properties include enhanced recombinogenicity, genome copy number, and capacity for expression and/or secretion of proteins and secondary metabolites.
1. A method of evolving a cell to acquire a desired functional property, the method comprising:
a) providing a population comprising genetically different haploid yeast cells of different mating type;
b) culturing the cells under conditions whereby the haploid yeast cells mate to form diploid cells;
c) culturing the mated diploid yeast cells under conditions whereby the cells recombine by meiosis to form haploid ascospores;
d) separating the haploid ascospores;
e) screening or selecting the separated haploid ascospores for cells that evolved toward acquisition of the desired functional property; and
f) repeating steps a) through e) wherein the recombined haploid cells identified in step e) are used as the population comprising genetically different yeast cells of different mating type provided in step a).
2. The method of claim 1 wherein the haploid ascospores are separated by degradation of the ascus with an enzyme.
3. The method of claim 2 wherein the enzyme is zymolase.
4. A method of evolving a cell to acquire a desired functional property, the method comprising:
a) providing a population comprising genetically different haploid yeast cells of different mating type;
b) culturing the cells under conditions whereby the haploid yeast cells mate to form diploid cells;
c) culturing the mated diploid yeast cells under conditions whereby the cells recombine by meiosis to form haploid ascospores;
d) separating the haploid ascospores;
e) repeating steps a) through d) whereby the separated haploid ascospores of step d) are used as the population comprising genetically different yeast cells of different mating type provided in step a); and
f) screening or selecting the separated haploid ascospores for cells that evolved toward acquisition of the desired functional property.
5. The method of claim 4 wherein the haploid ascospores are separated by degradation of the ascus with an enzyme.
6. The method of claim 5 wherein the enzyme is zymolase.