2. Academic Validation
  3. In vitro synthesis of gene-length single-stranded DNA

In vitro synthesis of gene-length single-stranded DNA

  • Sci Rep. 2018 Apr 25;8(1):6548. doi: 10.1038/s41598-018-24677-5.
Rémi Veneziano 1,  Tyson R Shepherd 2,  Sakul Ratanalert 2 3,  Leila Bellou 2,  Chaoqun Tao 2,  Mark Bathe 4
Affiliations

Affiliations

  • 1 Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA. [email protected].
  • 2 Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
  • 3 Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
  • 4 Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA. [email protected].
PMID: 29695837 DOI: 10.1038/s41598-018-24677-5
Abstract

Single-stranded DNA (ssDNA) increases the likelihood of homology directed repair with reduced cellular toxicity. However, ssDNA synthesis strategies are limited by the maximum length attainable, ranging from a few hundred nucleotides for chemical synthesis to a few thousand nucleotides for enzymatic synthesis, as well as limited control over nucleotide composition. Here, we apply purely enzymatic synthesis to generate ssDNA greater than 15 kilobases (kb) using asymmetric PCR, and illustrate the incorporation of diverse modified nucleotides for therapeutic and theranostic applications.

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