{
  "abstract": "Enzymatic synthesis of modified nucleic acids is gaining a lot of traction. Most approaches involve the combination of modified nucleoside triphosphates and engineered polymerases capable of accepting such altered substrates. Such enzymes are obtained through engineering campaigns based on directed evolution, positional saturation mutagenesis, and rational design approaches potentially guided by in silico methods. However, most polymerase evolution/design approaches rely on a restricted diversity of enzyme chassis. Here, we have performed a biochemical characterization of two polymerases, the human DNA HsPolθ and the heterodimeric archaeal DNA primase PriSL from Pyrococcus abyssi, capable of synthesizing oligonucleotides under both template-dependent and -independent reaction conditions. We have assessed their capacity at accepting nucleotides with modifications at any position of the scaffold as well as reversible terminators. We have also evaluated the possibility of using high salt conditions to force polymerases to incorporate single nucleotides. Overall, our study reveals that both polymerases are tolerant to modifications and could be amenable to engineering campaigns to identify variants capable of producing hypermodified XNAs for de novo nucleic acid synthesis approaches.",
  "abstract_shown": true,
  "affiliations": [
    "Department of Structural Biology and Chemistry, Laboratory for Bioorganic Chemistry of Nucleic Acids, 28, rue du Docteur Roux, Paris Cedex, France, 75724",
    "Chimie ParisTech, PSL University, CNRS, Institute of Chemistry for Life and Health Sciences, Laboratory for Inorganic Chemical Biology",
    "Institut Pasteur",
    "Unit of Architecture and Dynamics of Biological Macromolecules"
  ],
  "article_type": null,
  "arxiv": null,
  "author_total": 8,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Fabienne Levi-Acobas"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Germain Niogret"
    },
    {
      "affs": [
        0
      ],
      "name": "Sophie Battédou"
    },
    {
      "affs": [
        1
      ],
      "name": "Gilles Gasser"
    },
    {
      "affs": [
        2,
        3
      ],
      "name": "Markel Martinez Carranza"
    },
    {
      "affs": [
        2,
        3
      ],
      "name": "Marc Delarue"
    },
    {
      "affs": [
        2,
        3
      ],
      "name": "Ludovic Sauguet"
    },
    {
      "affs": [
        0
      ],
      "name": "Marcel Hollenstein",
      "orcid": "0000-0003-0263-9206"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/0rwpznfn13arc.bib",
  "cited_by": 0,
  "doi": "10.26434/chemrxiv.15009757/v1",
  "fulltext_url": null,
  "id": "0rwpznfn13arc",
  "issns": [],
  "landing_url": "https://chemrxiv.org/doi/full/10.26434/chemrxiv.15009757/v1",
  "language": null,
  "licence": "CC-BY",
  "licence_label": "CC BY",
  "licence_source": "journal",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "openalex",
      "1312|Enzyme Catalysis and Immobilization"
    ]
  ],
  "openalex": "W7215038576",
  "pdf_url": "https://chemrxiv.org/doi/pdf/10.26434/chemrxiv.15009757/v1",
  "pmcid": null,
  "pmid": null,
  "published": "2026-10-01",
  "publisher": "American Chemical Society (ACS)",
  "ris": "https://research.draconitic.com/paper/0rwpznfn13arc.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:17:21Z",
      "native_id": "10.26434/chemrxiv.15009757",
      "source": "chemrxiv",
      "url": "https://chemrxiv.org/doi/full/10.26434/chemrxiv.15009757/v1"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|Enzyme Catalysis and Immobilization",
      "subfield": "molecular-biology"
    }
  ],
  "title": "Evaluation of alternative polymerases for biocatalytic synthesis of modified oligonucleotides",
  "title_text": "Evaluation of alternative polymerases for biocatalytic synthesis of modified oligonucleotides",
  "type": "preprint",
  "updated_at": "2026-10-30T12:17:21Z",
  "url": "https://research.draconitic.com/paper/0rwpznfn13arc",
  "venue": "ChemRxiv",
  "versions": []
}