Modified template-independent dna polymerase
AssigneeThe Charles Stark Draper Laboratory, Inc.
InventorsAndrew P. Magyar, Peter Cavanagh, Miles Rogers, Kirsty A. McFarland, Melissa M. Sprachman, Amanda Nicole Billings-Siuti, Nicole E. Raustad
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Abstract
Described herein are genetically engineered template-independent DNA polymerases, specifically terminal deoxynucleotidyl transferases, and methods of using these polymerases to control DNA synthesis by adding a single nucleotide (mononucleotide) at a time to the 3′ end of a growing single-stranded DNA polynucleotide.
Claims44
An engineered terminal deoxynucleotidyl transferase (TdT), wherein one, or more, amino acid residues of the TdT are modified, resulting in a TdT capable of controlled addition of nucleotides to the 3′ end of a single-stranded polynucleotide.
A photoswitchable azobenzene moiety, wherein the azobenzene molecule is modified and the modification comprises introduction of an attachment site for a click reactive group and introduction of an attachment site for an amino acid side chain.
A kit comprising reagents for template-independent polynucleotide synthesis and a modified TdT of claim, wherein the TdT is capable of attaching a mononucleotide to the 3′ end of a single-stranded polynucleotide under conditions suitable for the incorporation of a mononucleotide to the 3′ end of the single-stranded polynucleotide.
Dependencies are inferred from the claim text.