Highly efficient genome editing in N. gerenzanensis using an inducible CRISPR/Cas9–RecA system
Objective To develop an inducible CRISPR/Cas9–Recombinase A (RecA) system to manipulate genes in Nonomuraea gerenzanensis effectively. Results Compared with traditional homologous recombination, the inducible CRISPR/Cas9 system achieved 68.8% editing efficiency, whereas, with both the inducible Cas9...
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| Published in | Biotechnology letters Vol. 42; no. 9; pp. 1699 - 1706 |
|---|---|
| Main Authors | , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Dordrecht
Springer Netherlands
01.09.2020
Springer Nature B.V |
| Subjects | |
| Online Access | Get full text |
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| Abstract | Objective
To develop an inducible CRISPR/Cas9–Recombinase A (RecA) system to manipulate genes in
Nonomuraea gerenzanensis
effectively.
Results
Compared with traditional homologous recombination, the inducible CRISPR/Cas9 system achieved 68.8% editing efficiency, whereas, with both the inducible Cas9 and the overexpressed RecA, the efficiency of the combined genome editing system reached 100%. The
dbv23
-deleted mutant obtained by the inducible CRISPR/Cas9–RecA system was confirmed to produce more A40926 with an approximate yield of 200 mg L
−1
than that of around 150 mg L
−1
produced by the wild-type strain.
Conclusions
This inducible CRISPR/Cas9–RecA system was successfully constructed and can be utilized as an efficient genome editing tool for
Actinomyce
s able to shorten editing time simultaneously. |
|---|---|
| AbstractList | OBJECTIVE: To develop an inducible CRISPR/Cas9–Recombinase A (RecA) system to manipulate genes in Nonomuraea gerenzanensis effectively. RESULTS: Compared with traditional homologous recombination, the inducible CRISPR/Cas9 system achieved 68.8% editing efficiency, whereas, with both the inducible Cas9 and the overexpressed RecA, the efficiency of the combined genome editing system reached 100%. The dbv23-deleted mutant obtained by the inducible CRISPR/Cas9–RecA system was confirmed to produce more A40926 with an approximate yield of 200 mg L⁻¹ than that of around 150 mg L⁻¹ produced by the wild-type strain. CONCLUSIONS: This inducible CRISPR/Cas9–RecA system was successfully constructed and can be utilized as an efficient genome editing tool for Actinomyces able to shorten editing time simultaneously. To develop an inducible CRISPR/Cas9-Recombinase A (RecA) system to manipulate genes in Nonomuraea gerenzanensis effectively. Compared with traditional homologous recombination, the inducible CRISPR/Cas9 system achieved 68.8% editing efficiency, whereas, with both the inducible Cas9 and the overexpressed RecA, the efficiency of the combined genome editing system reached 100%. The dbv23-deleted mutant obtained by the inducible CRISPR/Cas9-RecA system was confirmed to produce more A40926 with an approximate yield of 200 mg L than that of around 150 mg L produced by the wild-type strain. This inducible CRISPR/Cas9-RecA system was successfully constructed and can be utilized as an efficient genome editing tool for Actinomyces able to shorten editing time simultaneously. ObjectiveTo develop an inducible CRISPR/Cas9–Recombinase A (RecA) system to manipulate genes in Nonomuraea gerenzanensis effectively.ResultsCompared with traditional homologous recombination, the inducible CRISPR/Cas9 system achieved 68.8% editing efficiency, whereas, with both the inducible Cas9 and the overexpressed RecA, the efficiency of the combined genome editing system reached 100%. The dbv23-deleted mutant obtained by the inducible CRISPR/Cas9–RecA system was confirmed to produce more A40926 with an approximate yield of 200 mg L−1 than that of around 150 mg L−1 produced by the wild-type strain.ConclusionsThis inducible CRISPR/Cas9–RecA system was successfully constructed and can be utilized as an efficient genome editing tool for Actinomyces able to shorten editing time simultaneously. Objective To develop an inducible CRISPR/Cas9–Recombinase A (RecA) system to manipulate genes in Nonomuraea gerenzanensis effectively. Results Compared with traditional homologous recombination, the inducible CRISPR/Cas9 system achieved 68.8% editing efficiency, whereas, with both the inducible Cas9 and the overexpressed RecA, the efficiency of the combined genome editing system reached 100%. The dbv23 -deleted mutant obtained by the inducible CRISPR/Cas9–RecA system was confirmed to produce more A40926 with an approximate yield of 200 mg L −1 than that of around 150 mg L −1 produced by the wild-type strain. Conclusions This inducible CRISPR/Cas9–RecA system was successfully constructed and can be utilized as an efficient genome editing tool for Actinomyce s able to shorten editing time simultaneously. To develop an inducible CRISPR/Cas9-Recombinase A (RecA) system to manipulate genes in Nonomuraea gerenzanensis effectively.OBJECTIVETo develop an inducible CRISPR/Cas9-Recombinase A (RecA) system to manipulate genes in Nonomuraea gerenzanensis effectively.Compared with traditional homologous recombination, the inducible CRISPR/Cas9 system achieved 68.8% editing efficiency, whereas, with both the inducible Cas9 and the overexpressed RecA, the efficiency of the combined genome editing system reached 100%. The dbv23-deleted mutant obtained by the inducible CRISPR/Cas9-RecA system was confirmed to produce more A40926 with an approximate yield of 200 mg L-1 than that of around 150 mg L-1 produced by the wild-type strain.RESULTSCompared with traditional homologous recombination, the inducible CRISPR/Cas9 system achieved 68.8% editing efficiency, whereas, with both the inducible Cas9 and the overexpressed RecA, the efficiency of the combined genome editing system reached 100%. The dbv23-deleted mutant obtained by the inducible CRISPR/Cas9-RecA system was confirmed to produce more A40926 with an approximate yield of 200 mg L-1 than that of around 150 mg L-1 produced by the wild-type strain.This inducible CRISPR/Cas9-RecA system was successfully constructed and can be utilized as an efficient genome editing tool for Actinomyces able to shorten editing time simultaneously.CONCLUSIONSThis inducible CRISPR/Cas9-RecA system was successfully constructed and can be utilized as an efficient genome editing tool for Actinomyces able to shorten editing time simultaneously. |
| Author | Xia, Tianyu Li, Yongquan Yue, Xue Wang, Shuai Dong, Huijun |
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| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/32314149$$D View this record in MEDLINE/PubMed |
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| CitedBy_id | crossref_primary_10_1016_j_procbio_2021_01_007 crossref_primary_10_1186_s12934_022_01813_5 crossref_primary_10_3389_fbioe_2023_1335901 crossref_primary_10_1093_femsle_fnad006 crossref_primary_10_1021_acssynbio_4c00757 crossref_primary_10_1007_s10529_021_03210_1 crossref_primary_10_1007_s10529_023_03347_1 crossref_primary_10_1016_j_cbpa_2023_102367 crossref_primary_10_1021_acschembio_1c00170 crossref_primary_10_1021_acssynbio_1c00317 |
| Cites_doi | 10.1093/abbs/gmv007 10.1038/s41579-018-0002-7 10.1016/j.biocel.2019.105642 10.1093/nar/gkw660 10.1007/s00253-010-2579-2 10.3390/ijms19041089 10.1007/s10295-015-1682-x 10.1016/bs.mie.2014.10.036 10.1016/S1074-5521(03)00120-0 10.1038/ja.2009.127 10.1039/c4mt00318g 10.1126/sciadv.aav3335 10.1007/s10295-003-0024-6 10.1093/nar/gkw223 |
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| Snippet | Objective
To develop an inducible CRISPR/Cas9–Recombinase A (RecA) system to manipulate genes in
Nonomuraea gerenzanensis
effectively.
Results
Compared with... To develop an inducible CRISPR/Cas9-Recombinase A (RecA) system to manipulate genes in Nonomuraea gerenzanensis effectively. Compared with traditional... ObjectiveTo develop an inducible CRISPR/Cas9–Recombinase A (RecA) system to manipulate genes in Nonomuraea gerenzanensis effectively.ResultsCompared with... To develop an inducible CRISPR/Cas9-Recombinase A (RecA) system to manipulate genes in Nonomuraea gerenzanensis effectively.OBJECTIVETo develop an inducible... OBJECTIVE: To develop an inducible CRISPR/Cas9–Recombinase A (RecA) system to manipulate genes in Nonomuraea gerenzanensis effectively. RESULTS: Compared with... |
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| SubjectTerms | Actinobacteria - genetics Actinomyces Actinomyces - genetics Applied Microbiology Bacterial Proteins - genetics Biochemistry Biomedical and Life Sciences Biotechnology CRISPR CRISPR-Cas systems CRISPR-Cas Systems - genetics Editing Escherichia coli - genetics Gene Editing - methods Genome editing Genomes Homologous recombination Homology Life Sciences Microbiology mutants Mutation - genetics Nonomuraea Original Research Paper Rec A Recombinases - genetics RecA protein Recombinase |
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| Title | Highly efficient genome editing in N. gerenzanensis using an inducible CRISPR/Cas9–RecA system |
| URI | https://link.springer.com/article/10.1007/s10529-020-02893-2 https://www.ncbi.nlm.nih.gov/pubmed/32314149 https://www.proquest.com/docview/2428784148 https://www.proquest.com/docview/2393039983 https://www.proquest.com/docview/2551914038 |
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