In vitro synthesis of gene-length single-stranded DNA

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...

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Autores principales:	Rémi Veneziano, Tyson R. Shepherd, Sakul Ratanalert, Leila Bellou, Chaoqun Tao, Mark Bathe
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/cb86edf54d00480d9e19c68973dc90bb
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spelling	oai:doaj.org-article:cb86edf54d00480d9e19c68973dc90bb2021-12-02T15:08:37ZIn vitro synthesis of gene-length single-stranded DNA10.1038/s41598-018-24677-52045-2322https://doaj.org/article/cb86edf54d00480d9e19c68973dc90bb2018-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-24677-5https://doaj.org/toc/2045-2322Abstract 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.Rémi VenezianoTyson R. ShepherdSakul RatanalertLeila BellouChaoqun TaoMark BatheNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-7 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Rémi Veneziano Tyson R. Shepherd Sakul Ratanalert Leila Bellou Chaoqun Tao Mark Bathe In vitro synthesis of gene-length single-stranded DNA
description	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.
format	article
author	Rémi Veneziano Tyson R. Shepherd Sakul Ratanalert Leila Bellou Chaoqun Tao Mark Bathe
author_facet	Rémi Veneziano Tyson R. Shepherd Sakul Ratanalert Leila Bellou Chaoqun Tao Mark Bathe
author_sort	Rémi Veneziano
title	In vitro synthesis of gene-length single-stranded DNA
title_short	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_sort	in vitro synthesis of gene-length single-stranded dna
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/cb86edf54d00480d9e19c68973dc90bb
work_keys_str_mv	AT remiveneziano invitrosynthesisofgenelengthsinglestrandeddna AT tysonrshepherd invitrosynthesisofgenelengthsinglestrandeddna AT sakulratanalert invitrosynthesisofgenelengthsinglestrandeddna AT leilabellou invitrosynthesisofgenelengthsinglestrandeddna AT chaoquntao invitrosynthesisofgenelengthsinglestrandeddna AT markbathe invitrosynthesisofgenelengthsinglestrandeddna
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In vitro synthesis of gene-length single-stranded DNA

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