Genetic editing of the virulence gene of Escherichia coli using the CRISPR system
Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 is an emerging gene-editing technology that is widely used in prokaryotes and eukaryotes. It can realize the specific manipulation of the genome efficiently and accurately. CRISPR/Cas9 coupled λ-Red recombination technology was...
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PeerJ Inc.
2020-04-01
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| Online Access: | https://peerj.com/articles/8881.pdf |
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| author | Meijia Hou Simeng Sun Qizheng Feng Xiumei Dong Ping Zhang Bo Shi Jiali Liu Dongfang Shi |
| author_facet | Meijia Hou Simeng Sun Qizheng Feng Xiumei Dong Ping Zhang Bo Shi Jiali Liu Dongfang Shi |
| author_sort | Meijia Hou |
| collection | DOAJ |
| description | Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 is an emerging gene-editing technology that is widely used in prokaryotes and eukaryotes. It can realize the specific manipulation of the genome efficiently and accurately. CRISPR/Cas9 coupled λ-Red recombination technology was used to perform genome editing in different genes. For finding an efficient method to edit the virulence genes of enterotoxigenic E. coli (ETEC), the two-plasmid system was used. The coding sequence (CDS) region of the estA, eltI, estB, eltIIc1, and faeG locus were deleted. The coding region of estB was substituted with estA. Gene recombination efficiency ranged from 0 to 77.78% when the length of the homology arm was from 50 to 300 bp. Within this range, the longer the homology arm, the higher the efficiency of genetic recombination. The results showed that this system can target virulence genes located in plasmids and on chromosomes of ETEC strains. A single base mutation was performed by two-step gene fragment replacement. This study lays the foundation for research on virulence factors and genetic engineering of vaccines for ETEC. |
| first_indexed | 2024-03-09T08:06:44Z |
| format | Article |
| id | doaj.art-a1c41069aba64764807597c4d58208c1 |
| institution | Directory Open Access Journal |
| issn | 2167-8359 |
| language | English |
| last_indexed | 2024-03-09T08:06:44Z |
| publishDate | 2020-04-01 |
| publisher | PeerJ Inc. |
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| spelling | doaj.art-a1c41069aba64764807597c4d58208c12023-12-02T23:48:24ZengPeerJ Inc.PeerJ2167-83592020-04-018e888110.7717/peerj.8881Genetic editing of the virulence gene of Escherichia coli using the CRISPR systemMeijia Hou0Simeng Sun1Qizheng Feng2Xiumei Dong3Ping Zhang4Bo Shi5Jiali Liu6Dongfang Shi7Veterinary Medicine, Northeast Agricultural University, Harbin, Heilongjiang Province, ChinaVeterinary Medicine, Northeast Agricultural University, Harbin, Heilongjiang Province, ChinaVeterinary Medicine, Northeast Agricultural University, Harbin, Heilongjiang Province, ChinaVeterinary Medicine, Northeast Agricultural University, Harbin, Heilongjiang Province, ChinaVeterinary Medicine, Northeast Agricultural University, Harbin, Heilongjiang Province, ChinaVeterinary Medicine, Northeast Agricultural University, Harbin, Heilongjiang Province, ChinaVeterinary Medicine, Northeast Agricultural University, Harbin, Heilongjiang Province, ChinaVeterinary Medicine, Northeast Agricultural University, Harbin, Heilongjiang Province, ChinaClustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 is an emerging gene-editing technology that is widely used in prokaryotes and eukaryotes. It can realize the specific manipulation of the genome efficiently and accurately. CRISPR/Cas9 coupled λ-Red recombination technology was used to perform genome editing in different genes. For finding an efficient method to edit the virulence genes of enterotoxigenic E. coli (ETEC), the two-plasmid system was used. The coding sequence (CDS) region of the estA, eltI, estB, eltIIc1, and faeG locus were deleted. The coding region of estB was substituted with estA. Gene recombination efficiency ranged from 0 to 77.78% when the length of the homology arm was from 50 to 300 bp. Within this range, the longer the homology arm, the higher the efficiency of genetic recombination. The results showed that this system can target virulence genes located in plasmids and on chromosomes of ETEC strains. A single base mutation was performed by two-step gene fragment replacement. This study lays the foundation for research on virulence factors and genetic engineering of vaccines for ETEC.https://peerj.com/articles/8881.pdfCRISPR-Cas9Virulence genesETECSingle base mutationCDS |
| spellingShingle | Meijia Hou Simeng Sun Qizheng Feng Xiumei Dong Ping Zhang Bo Shi Jiali Liu Dongfang Shi Genetic editing of the virulence gene of Escherichia coli using the CRISPR system PeerJ CRISPR-Cas9 Virulence genes ETEC Single base mutation CDS |
| title | Genetic editing of the virulence gene of Escherichia coli using the CRISPR system |
| title_full | Genetic editing of the virulence gene of Escherichia coli using the CRISPR system |
| title_fullStr | Genetic editing of the virulence gene of Escherichia coli using the CRISPR system |
| title_full_unstemmed | Genetic editing of the virulence gene of Escherichia coli using the CRISPR system |
| title_short | Genetic editing of the virulence gene of Escherichia coli using the CRISPR system |
| title_sort | genetic editing of the virulence gene of escherichia coli using the crispr system |
| topic | CRISPR-Cas9 Virulence genes ETEC Single base mutation CDS |
| url | https://peerj.com/articles/8881.pdf |
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