FIND Technology
Encyclopedia
FIND® technology is a directed evolution
technology that uses DNA recombination to improve properties of proteins. It eliminates unimportant and deleterious mutations while maintaining and combining beneficial mutations that would enhance protein function.
. The newly mutated library is then subjected to exonuclease activity, creating both sense and anti-sense fragments. The areas of partially overlapping fragments are then combined and extended using a PCR-like method. These double stranded mutants are then screened for the desired optimized function using a relevant assay. The best mutants are chosen for further exonuclease
activity. The process (exonuclease, PCR-like recombination, and mutant screening) is repeated, usually about 10-12 times, in order to achieve the best possible mutants with only beneficial mutations.
Directed evolution
thumb|250px|right|An example of a possible round to evolve a protein based fluorescent sensor for a specific analyte using two consecutive FACS sortings...
technology that uses DNA recombination to improve properties of proteins. It eliminates unimportant and deleterious mutations while maintaining and combining beneficial mutations that would enhance protein function.
Procedures
For the relevant gene, a library of single stranded oligonucleotides is acquired and then mutated using random mutagenesisMutagenesis
Mutagenesis is a process by which the genetic information of an organism is changed in a stable manner, resulting in a mutation. It may occur spontaneously in nature, or as a result of exposure to mutagens. It can also be achieved experimentally using laboratory procedures...
. The newly mutated library is then subjected to exonuclease activity, creating both sense and anti-sense fragments. The areas of partially overlapping fragments are then combined and extended using a PCR-like method. These double stranded mutants are then screened for the desired optimized function using a relevant assay. The best mutants are chosen for further exonuclease
Exonuclease
Exonucleases are enzymes that work by cleaving nucleotides one at a time from the end of a polynucleotide chain. A hydrolyzing reaction that breaks phosphodiester bonds at either the 3’ or the 5’ end occurs. Its close relative is the endonuclease, which cleaves phosphodiester bonds in the middle ...
activity. The process (exonuclease, PCR-like recombination, and mutant screening) is repeated, usually about 10-12 times, in order to achieve the best possible mutants with only beneficial mutations.