US 12,071,638 B2
Variants of terminal deoxynucleotidyl transferase and uses thereof
Elise Champion, Paris (FR); Mikhael Soskine, Franconville (FR); Thomas Ybert, Paris (FR); and Marc Delarue, Versailles (FR)
Assigned to DNA Script, Le Kremlin-Bicêtre (FR); Institut Pasteur, Le Kremlin-Bicêtre (FR); and Centre National de la Recherche Scientifique, Paris (FR)
Filed by DNA Script, Paris (FR); Institut Pasteur, Paris (FR); and Centre National de la Recherche Scientifique, Paris (FR)
Filed on Dec. 7, 2021, as Appl. No. 17/544,647.
Application 17/544,647 is a continuation of application No. 16/925,785, filed on Jul. 10, 2020, granted, now 11,208,637.
Application 16/925,785 is a continuation of application No. 16/423,972, filed on May 28, 2019, granted, now 10,752,887, issued on Aug. 25, 2020.
Application 16/423,972 is a continuation in part of application No. 16/242,904, filed on Jan. 8, 2019, granted, now 10,435,676, issued on Oct. 8, 2019.
Claims priority of application No. 18305006 (EP), filed on Jan. 8, 2018.
Prior Publication US 2022/0403354 A1, Dec. 22, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/70 (2006.01); C12N 9/12 (2006.01)
CPC C12N 9/1264 (2013.01) [C12N 15/70 (2013.01); C12Y 207/07031 (2013.01)] 22 Claims
 
1. A terminal deoxynucleotidyl transferase (TdT) variant comprising an amino acid sequence at least 90% identical to SEQ ID NO:2, SEQ ID NO:11, SEQ ID NO: 13, SEQ ID NO: 17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:29, or SEQ ID NO:31 with an amino acid substitution of cysteine at position 173, wherein the TdT variant (i) is capable of synthesizing a nucleic acid fragment without a template and (ii) is capable of incorporating a 3′-O-modified nucleotide into the nucleic acid fragment, and
wherein the TdT variant incorporates the 3′-O-modified nucleotide at a rate greater than that of a wild type TdT.