Abstract: The present disclosure provides modified terminal deoxynucleotidyl (TdT) polymerases and compositions (e.g., conjugates) comprising modified TdT polymerases. The present disclosure also provides circularly permuted terminal deoxynucleotidyl (TdT) polymerases as well as modified TdT polymerases and compositions (e.g., conjugates) comprising such cpTdT or modified TdT polymerases. Also provided are nucleic acids, vectors, and host cells encoding modified TdT polymerases, as well as methods including methods of producing modified TdT polymerases, and methods of using modified TdT polymerases (e.g., methods of nucleic acid synthesis).

Abstract: Disclosed herein are compositions and methods for de novo enzymatic synthesis of nucleic acid molecules. In particular, and among other things, compositions and methods of the present disclosure provide for reduction of non-terminations in a nucleic acid molecule product generated by de novo enzymatic synthesis. Compositions and methods of the present disclosure also include the use of a phosphatase enzyme to facilitate the reduction of non-terminations.

Abstract: Methods and compositions related to intracellular delivery of gene editing proteins are provided. The invention relates to compositions and methods for transporting gene editing polypeptides, such as Cas9 or Cas12, into a cell ex vivo or in vivo. The invention includes a targeted active gene editing (TAGE) agent that includes an extracellular cell membrane binding moiety, e.g., an antigen binding polypeptide, a cell penetrating peptide (CPP), a ligand, or combinations thereof, that specifically binds to an extracellular cell membrane-bound molecule (e.g., a cell surface molecule), and a site-directed modifying polypeptide that recognizes a nucleic acid sequence. The extracellular cell membrane binding moiety (e.g.

Abstract: Methods and compositions related to intracellular delivery of gene editing proteins are provided. The invention relates to compositions and methods for transporting gene editing polypeptides, such as Cas9 or Cas12, into a cell ex vivo or in vivo. The invention includes a targeted active gene editing (TAGE) agent that includes an antigen binding polypeptide that specifically binds to an extracellular cell membrane-bound molecule, and a site-directed modifying polypeptide that recognizes a nucleic acid sequence. The antigen binding polypeptide and the site-directed modifying polypeptide are stably associated such that the site-directed modifying polypeptide can be internalized into a cell displaying the extracellular cell membrane-bound molecule.