中文版 | English
Title

Customized one-step preparation of sgRNA transcription templates via overlapping PCR Using short primers and its application in vitro and in vivo gene editing

Author
Publication Years
2019-10-24
Keywords
Language
English
URL[Source Record]
Abstract
Overlap extension polymerase chain reaction (PCR) is a powerful technology for DNA assembly. Based on this technology, we synthesized DNA templates, which were transcribed into sgRNA in vitro, and further detected their efficiency of purified sgRNAs with Cas9 nuclease. The sgRNAs synthesized by this approach can effectively cleave the DNA fragments of interest in vitro and in vivo. Compared with the conventional method for generating sgRNA, it does not require construction of recombinant plasmids and design of primers to amplify sgRNA core fragment. Only several short primers with overlapped sequences are needed to assemble a DNA fragment as the template of sgRNA. This modified and simplified method is highly applicable and less time-consuming.
DOI
Source Title
Volume
9
Issue
1
Indexed By
SUSTech Authorship
Others
Scopus EID
2-s2.0-85074460542
Data Source
Scopus
Corresponding AuthorHu,Zheng
WOS Accession No
WOS:000492859400001
EISSN
2045-3701
Citation statistics
Cited Times [WOS]:7
Document TypeOther
Identifierhttp://kc.sustech.edu.cn/handle/2SGJ60CL/407164
DepartmentDepartment of Biology
生命科学学院
南方科技大学医学院
Affiliation
1.First People's Hospital of Chenzhou,Affiliated to University of Southern Medical University,Chenzhou,423000,China
2.National and Local Joint Engineering Laboratory for High-throughput Molecular Diagnosis Technology,Affiliated to the First People's Hospital of Chenzhou,University of South China,Chenzhou,423000,China
3.Translational Medicine Institute,University of South China,Chenzhou,423000,China
4.Department of Biology,Guangdong Provincial Key Laboratory of Cell Microenvironment and Disease Research,Shenzhen Key Laboratory of Cell Microenvironment,Southern University of Science and Technology,Shenzhen, Guangdong,518055,China
5.Genetalks Bio-tech (Changsha) Limited Liability Company,Changsha, Hunan,410013,China
Recommended Citation
GB/T 7714
Hu,Zheng,Wang,Li,Shi,Zhaoying,et al. Customized one-step preparation of sgRNA transcription templates via overlapping PCR Using short primers and its application in vitro and in vivo gene editing. 2019-10-24.
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