In vitro synthesis of gene-length single-stranded DNA
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...
| Main Authors: | , , , , , |
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| Format: | Article |
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Nature Publishing Group
2018
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| Online Access: | http://hdl.handle.net/1721.1/117554 https://orcid.org/0000-0002-2726-3770 https://orcid.org/0000-0001-7122-1917 https://orcid.org/0000-0002-1766-807X https://orcid.org/0000-0002-6199-6855 |
| _version_ | 1811087597000196096 |
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| author | Veneziano, Remi Shepherd, Tyson R Ratanalert, Sakul Bellou, Leila Tao, Chaoqun Bathe, Mark |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Veneziano, Remi Shepherd, Tyson R Ratanalert, Sakul Bellou, Leila Tao, Chaoqun Bathe, Mark |
| author_sort | Veneziano, Remi |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T13:48:37Z |
| format | Article |
| id | mit-1721.1/117554 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T13:48:37Z |
| publishDate | 2018 |
| publisher | Nature Publishing Group |
| record_format | dspace |
| spelling | mit-1721.1/1175542022-10-01T17:18:31Z In vitro synthesis of gene-length single-stranded DNA Veneziano, Remi Shepherd, Tyson R Ratanalert, Sakul Bellou, Leila Tao, Chaoqun Bathe, Mark Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Chemical Engineering Veneziano, Remi Shepherd, Tyson R Ratanalert, Sakul Bellou, Leila Tao, Chaoqun Bathe, Mark 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. United States. Office of Naval Research (Grant N00014-17-1-2609) United States. Office of Naval Research (Grant N00014-16-1-2181) United States. Office of Naval Research (Grant N00014-16-1-2953) National Science Foundation (U.S.) (Grant EAGER CCF-1547999) 2018-08-27T18:20:22Z 2018-08-27T18:20:22Z 2018-04 2017-12 2018-08-27T12:51:41Z Article http://purl.org/eprint/type/JournalArticle 2045-2322 http://hdl.handle.net/1721.1/117554 Veneziano, Rémi et al. “In Vitro Synthesis of Gene-Length Single-Stranded DNA.” Scientific Reports 8, 1 (April 2018): 6548 © 2018 The Author(s). https://orcid.org/0000-0002-2726-3770 https://orcid.org/0000-0001-7122-1917 https://orcid.org/0000-0002-1766-807X https://orcid.org/0000-0002-6199-6855 http://dx.doi.org/10.1038/s41598-018-24677-5 Scientific Reports Creative Commons Attribution 4.0 International License http://creativecommons.org/licenses/by/4.0/ application/pdf Nature Publishing Group Nature |
| spellingShingle | Veneziano, Remi Shepherd, Tyson R Ratanalert, Sakul Bellou, Leila Tao, Chaoqun Bathe, Mark In vitro synthesis of gene-length single-stranded DNA |
| title | In vitro synthesis of gene-length single-stranded DNA |
| title_full | In vitro synthesis of gene-length single-stranded DNA |
| title_fullStr | In vitro synthesis of gene-length single-stranded DNA |
| title_full_unstemmed | In vitro synthesis of gene-length single-stranded DNA |
| title_short | In vitro synthesis of gene-length single-stranded DNA |
| title_sort | in vitro synthesis of gene length single stranded dna |
| url | http://hdl.handle.net/1721.1/117554 https://orcid.org/0000-0002-2726-3770 https://orcid.org/0000-0001-7122-1917 https://orcid.org/0000-0002-1766-807X https://orcid.org/0000-0002-6199-6855 |
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