In vitro synthesis of gene-length single-stranded DNA
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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.
Date issued
2018-04Department
Massachusetts Institute of Technology. Department of Biological Engineering; Massachusetts Institute of Technology. Department of Chemical EngineeringJournal
Scientific Reports
Publisher
Nature Publishing Group
Citation
Veneziano, Rémi et al. “In Vitro Synthesis of Gene-Length Single-Stranded DNA.” Scientific Reports 8, 1 (April 2018): 6548 © 2018 The Author(s).
Version: Final published version
ISSN
2045-2322