Highly efficient editing of the actinorhodin polyketide chain length factor gene in Streptomyces coelicolor M145 using CRISPR/Cas9-CodA(sm) combined system

Verfasser / Beitragende:
[Hu Zeng, Shishi Wen, Wei Xu, Zhaoren He, Guifa Zhai, Yunkun Liu, Zixin Deng, Yuhui Sun]
Ort, Verlag, Jahr:
2015
Enthalten in:
Applied Microbiology and Biotechnology, 99/24(2015-12-01), 10575-10585
Format:
Artikel (online)
ID: 605502676
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024 7 0 |a 10.1007/s00253-015-6931-4  |2 doi 
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245 0 0 |a Highly efficient editing of the actinorhodin polyketide chain length factor gene in Streptomyces coelicolor M145 using CRISPR/Cas9-CodA(sm) combined system  |h [Elektronische Daten]  |c [Hu Zeng, Shishi Wen, Wei Xu, Zhaoren He, Guifa Zhai, Yunkun Liu, Zixin Deng, Yuhui Sun] 
520 3 |a The current diminishing returns in finding useful antibiotics and the occurrence of drug-resistant bacteria call for the need to find new antibiotics. Moreover, the whole genome sequencing revealed that the biosynthetic potential of Streptomyces, which has produced the highest numbers of approved and clinical-trial drugs, has been greatly underestimated. Considering the known gene editing toolkits were arduous and inefficient, novel and efficient gene editing system are desirable. Here, we developed an engineered CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat/CRISPR-associated protein) combined with the counterselection system CodA(sm), the D314A mutant of cytosine deaminase, to rapidly and effectively edit Streptomyces genomes. In-frame deletion of the actinorhodin polyketide chain length factor gene actI-ORF2 was created in Streptomyces coelicolor M145 as an illustration. This CRISPR/Cas9-CodA(sm) combined system strikingly increased the frequency of unmarked mutants and shortened the time required to generate them. We foresee the system becoming a routine laboratory technique for genome editing to exploit the great biosynthetic potential of Streptomyces and perhaps for other medically and economically important actinomycetes. 
540 |a Springer-Verlag Berlin Heidelberg, 2015 
690 7 |a CRISPR/Cas9  |2 nationallicence 
690 7 |a CodA  |2 nationallicence 
690 7 |a Actinorhodin biosynthesis  |2 nationallicence 
690 7 |a ActI-ORF2  |2 nationallicence 
690 7 |a Gene editing  |2 nationallicence 
700 1 |a Zeng  |D Hu  |u Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education and Wuhan University School of Pharmaceutical Sciences, 185 East Lake Road, 430071, Wuhan, People's Republic of China  |4 aut 
700 1 |a Wen  |D Shishi  |u Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education and Wuhan University School of Pharmaceutical Sciences, 185 East Lake Road, 430071, Wuhan, People's Republic of China  |4 aut 
700 1 |a Xu  |D Wei  |u Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education and Wuhan University School of Pharmaceutical Sciences, 185 East Lake Road, 430071, Wuhan, People's Republic of China  |4 aut 
700 1 |a He  |D Zhaoren  |u Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education and Wuhan University School of Pharmaceutical Sciences, 185 East Lake Road, 430071, Wuhan, People's Republic of China  |4 aut 
700 1 |a Zhai  |D Guifa  |u Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education and Wuhan University School of Pharmaceutical Sciences, 185 East Lake Road, 430071, Wuhan, People's Republic of China  |4 aut 
700 1 |a Liu  |D Yunkun  |u Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education and Wuhan University School of Pharmaceutical Sciences, 185 East Lake Road, 430071, Wuhan, People's Republic of China  |4 aut 
700 1 |a Deng  |D Zixin  |u Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education and Wuhan University School of Pharmaceutical Sciences, 185 East Lake Road, 430071, Wuhan, People's Republic of China  |4 aut 
700 1 |a Sun  |D Yuhui  |u Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education and Wuhan University School of Pharmaceutical Sciences, 185 East Lake Road, 430071, Wuhan, People's Republic of China  |4 aut 
773 0 |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/24(2015-12-01), 10575-10585  |x 0175-7598  |q 99:24<10575  |1 2015  |2 99  |o 253 
856 4 0 |u https://doi.org/10.1007/s00253-015-6931-4  |q text/html  |z Onlinezugriff via DOI 
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950 |B NATIONALLICENCE  |P 856  |E 40  |u https://doi.org/10.1007/s00253-015-6931-4  |q text/html  |z Onlinezugriff via DOI 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Zeng  |D Hu  |u Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education and Wuhan University School of Pharmaceutical Sciences, 185 East Lake Road, 430071, Wuhan, People's Republic of China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Wen  |D Shishi  |u Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education and Wuhan University School of Pharmaceutical Sciences, 185 East Lake Road, 430071, Wuhan, People's Republic of China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Xu  |D Wei  |u Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education and Wuhan University School of Pharmaceutical Sciences, 185 East Lake Road, 430071, Wuhan, People's Republic of China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a He  |D Zhaoren  |u Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education and Wuhan University School of Pharmaceutical Sciences, 185 East Lake Road, 430071, Wuhan, People's Republic of China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Zhai  |D Guifa  |u Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education and Wuhan University School of Pharmaceutical Sciences, 185 East Lake Road, 430071, Wuhan, People's Republic of China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Liu  |D Yunkun  |u Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education and Wuhan University School of Pharmaceutical Sciences, 185 East Lake Road, 430071, Wuhan, People's Republic of China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Deng  |D Zixin  |u Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education and Wuhan University School of Pharmaceutical Sciences, 185 East Lake Road, 430071, Wuhan, People's Republic of China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Sun  |D Yuhui  |u Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education and Wuhan University School of Pharmaceutical Sciences, 185 East Lake Road, 430071, Wuhan, People's Republic of China  |4 aut 
950 |B NATIONALLICENCE  |P 773  |E 0-  |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/24(2015-12-01), 10575-10585  |x 0175-7598  |q 99:24<10575  |1 2015  |2 99  |o 253