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...

Descripción completa

Detalles Bibliográficos
Autores principales: Veneziano, Rémi, Shepherd, Tyson R., Ratanalert, Sakul, Bellou, Leila, Tao, Chaoqun, Bathe, Mark
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5916881/
https://www.ncbi.nlm.nih.gov/pubmed/29695837
http://dx.doi.org/10.1038/s41598-018-24677-5
_version_ 1783317081557565440
author Veneziano, Rémi
Shepherd, Tyson R.
Ratanalert, Sakul
Bellou, Leila
Tao, Chaoqun
Bathe, Mark
author_facet Veneziano, Rémi
Shepherd, Tyson R.
Ratanalert, Sakul
Bellou, Leila
Tao, Chaoqun
Bathe, Mark
author_sort Veneziano, Rémi
collection PubMed
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.
format Online
Article
Text
id pubmed-5916881
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-59168812018-04-30 In vitro synthesis of gene-length single-stranded DNA Veneziano, Rémi Shepherd, Tyson R. Ratanalert, Sakul Bellou, Leila Tao, Chaoqun Bathe, Mark Sci Rep Article 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. Nature Publishing Group UK 2018-04-25 /pmc/articles/PMC5916881/ /pubmed/29695837 http://dx.doi.org/10.1038/s41598-018-24677-5 Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Veneziano, Rémi
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
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5916881/
https://www.ncbi.nlm.nih.gov/pubmed/29695837
http://dx.doi.org/10.1038/s41598-018-24677-5
work_keys_str_mv AT venezianoremi invitrosynthesisofgenelengthsinglestrandeddna
AT shepherdtysonr invitrosynthesisofgenelengthsinglestrandeddna
AT ratanalertsakul invitrosynthesisofgenelengthsinglestrandeddna
AT bellouleila invitrosynthesisofgenelengthsinglestrandeddna
AT taochaoqun invitrosynthesisofgenelengthsinglestrandeddna
AT bathemark invitrosynthesisofgenelengthsinglestrandeddna

Ejemplares similares