Evolving new molecular function

An in vitro method of performing synthesis from a nucleic acid template to produce a reaction product, wherein the reaction product is not a nucleic acid, the method comprising the steps of:

• (a) providing (i) an amplifiable template comprising a first reactive unit associated with a first oligonucleotide defining a first codon sequence, and (ii) a first transfer unit comprising a second reactive unit associated with a second oligonucleotide defining an anti-codon sequence complementary to the first codon sequence;
• (b) annealing the codon and anti-codon sequences to bring the first reactive unit and the second reactive unit into reactive proximity;
• (c) inducing a reaction between the first and second reactive units without assistance of a ribosome to produce the reaction product;
• and
• (d) amplifying the template that encoded the reaction product.

An in vitro method of performing synthesis from a nucleic acid template to produce a reaction product, wherein the reaction product is not a nucleic acid, the method comprising the steps of:

• (a) providing (i) a template comprising a first reactive unit associated with a first oligonucleotide defining a first codon sequence and a second, different codon sequence, and (ii) a first transfer unit comprising a second reactive unit associated with a second oligonucleotide defining a first anti-codon sequence complementary to the first codon sequence of the template;
• (b) annealing the first codon sequence and the first anti-codon sequence to bring the first reactive unit and the second reactive unit into reactive proximity;
• (c) inducing a reaction between the first and second reactive units without assistance of a ribosome to produce the reaction product;
• and
• (d) selecting the reaction product still associated with the template.

An in vitro method of performing synthesis from a nucleic acid template to produce a modified scaffold molecule, the method comprising the steps of:

• (a) providing (i) a template comprising a scaffold molecule associated with a first oligonucleotide defining a first codon sequence and a second, different codon sequence, the scaffold molecule comprising a plurality of sites for modification, and (ii) a first transfer unit comprising a reactive unit covalently attached to a second oligonucleotide defining a first anti-codon sequence complementary to the first codon sequence;
• (b) annealing the first codon sequence and the first anti-codon sequence to bring the scaffold molecule and the reactive unit into reactive proximity;
• and
• (c) inducing a covalent bond-forming reaction between the reactive unit and the scaffold molecule without assistance of a ribosome to produce the modified scaffold molecule.

An in vitro method of producing a plurality of different reaction products in a single solution, the method comprising the steps of:

• (a) providing a plurality of different templates comprising a plurality of first reactive units associated with a corresponding plurality of first oligonucleotides, wherein each first oligonucleotide defines at least one codon sequence;
• (b) providing a plurality of transfer units comprising a plurality of second reactive units covalently attached to a corresponding plurality of second oligonucleotides, wherein the second oligonueleotides each define an anti-codon sequence complementary to a codon sequence;
• (c) annealing oligonucleotides having complementary codon and anti-codon sequences to bring the first and second reactive units into reactive proximity;
• and
• (d) inducing covalent bond-forming reactions between the reactive units without the assistance of a ribosome to produce the plurality of different reaction products.