In vitro synthesis of gene-length single-stranded DNA

被引：62

作者：

Veneziano, Remi ^{[1
]}

Shepherd, Tyson R. ^{[1
]}

Ratanalert, Sakul ^{[1
,2
]}

Bellou, Leila ^{[1
]}

Tao, Chaoqun ^{[1
]}

Bathe, Mark ^{[1
]}

机构：

[1] MIT, Dept Biol Engn, 77 Massachusetts Ave, Cambridge, MA 02139 USA

[2] MIT, Dept Chem Engn, Cambridge, MA 02139 USA

来源：

SCIENTIFIC REPORTS | 2018年 / 8卷

关键词：

ASYMMETRIC PCR; STABILITY; MOLECULES; IMAGE;

D O I：

10.1038/s41598-018-24677-5

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页数：7

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