Double-Check Base Editing for Efficient A to G Conversions
| Autor: | Siwei Li, Feiyu Fan, Zhongkang Li, Qianwen Xie, Xueli Zhang, Changhao Bi, Ju Li, Dongdong Zhao, Xiuqing Xin |
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| Rok vydání: | 2019 |
| Předmět: |
0106 biological sciences
Computer science Guanine Biomedical Engineering Computational biology 01 natural sciences Biochemistry Genetics and Molecular Biology (miscellaneous) 03 medical and health sciences chemistry.chemical_compound Cytosine Adenosine deaminase 010608 biotechnology CRISPR-Associated Protein 9 Escherichia coli CRISPR Double check 030304 developmental biology Gene Editing 0303 health sciences biology Base Sequence Cas9 APOBEC1 Adenine General Medicine Cytidine deaminase chemistry biology.protein Plasmids |
| Zdroj: | ACS synthetic biology. 8(12) |
| ISSN: | 2161-5063 |
| Popis: | With the development of CRISPR/Cas9 technology, a new generation of editing methods that convert specific bases has enabled precise single-base mutations. To date, conversion of cytosine to thymidine and adenine to guanine has been achieved using the cytidine deaminase APOBEC1 and adenosine deaminase (TadA), respectively. However, the base editing efficiency can be unacceptably low in some cell types or at certain target loci. One reason might be the lack of a selective pressure against the survival of nonedited cells. Few studies on ABE in prokaryotes have been reported, probably due to the relatively low editing efficiency of TadA. Improving the editing efficiency is the key for establishing base editing techniques and especially the ABE technologies. In this work, a selective pressure against nonedited cells was implemented to increase the base editing efficiency. First, we fused nCas9 or dCas9 with TadA to compare the editing efficiency of nCas9-TadA and dCas9-TadA fusion complexes in the model prokar... |
| Databáze: | OpenAIRE |
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