About: A programmable CRISPR/Cas9-based phage defense system for Escherichia coli BL21(DE3)

Facets (new session)
Description
Metadata
Settings
- owl:sameAs
- Inference Rule:

About: A programmable CRISPR/Cas9-based phage defense system for Escherichia coli BL21(DE3) Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : covidontheweb.inria.fr associated with source document(s)

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	A programmable CRISPR/Cas9-based phage defense system for Escherichia coli BL21(DE3)
Creator	Liu, Li Zhang, Xueli Bi, Changhao Hu, Muzi Xiong, Bin Ye, Lijun Zhan, Tao Zhao, Dongdong
Source	Medline; PMC
abstract	Escherichia coli BL21 is arguably the most popular host for industrial production of proteins, and industrial fermentations are often plagued by phage infections. The CRISPR/Cas system is guided by a gRNA to cleave a specific DNA cassette, which can be developed into a highly efficient programable phage defense system. In this work, we constructed a CRISPR/Cas system targeting multiple positions on the genome of T7 phage and found that the system increased the BL21’s defense ability against phage infection. Furthermore, the targeted loci on phage genome played a critical role. For better control of expression of CRISPR/Cas9, various modes were tested, and the OD of the optimized strain BL21(pT7cas9, pT7-3gRNA, prfp) after 4 h of phage infection was significantly improved, reaching 2.0, which was similar to the control culture without phage infection. Although at later time points, the defensive ability of CRISPR/Cas9 systems were not as obvious as that at early time points. The viable cell count of the engineered strain in the presence of phage was only one order of magnitude lower than that of the strain with no infection, which further demonstrated the effectiveness of the CRISPR/Cas9 phage defense system. Finally, the engineered BL21 strain under phage attack expressed RFP protein at about 60% of the un-infected control, which was significantly higher than the parent BL21. In this work, we successfully constructed a programable CRISPR/Cas9 system to increase the ability of E. coli BL21’s to defend against phage infection, and created a resistant protein expression host. This work provides a simple and feasible strategy for protecting industrial E. coli strains against phage infection.
has issue date	2020-07-03(xsd:dateTime)
bibo:doi	10.1186/s12934-020-01393-2
bibo:pmid	32620105
has license	cc-by
sha1sum (hex)	dee99e26e83d0048ad301eefc8438f26db0f54c8
schema:url	https://doi.org/10.1186/s12934-020-01393-2
resource representing a document's title	A programmable CRISPR/Cas9-based phage defense system for Escherichia coli BL21(DE3)
has PubMed Central identifier	PMC7332536
has PubMed identifier	32620105
schema:publication	Microb Cell Fact
resource representing a document's body	covid:dee99e26e83d0048ad301eefc8438f26db0f54c8#body_text
is schema:about of	named entity 'phage' named entity 'infection' named entity 'proteins' named entity 'viable' named entity 'tested' named entity 'constructed' named entity 'CRISPR/Cas9' named entity 'system' named entity 'attack' named entity 'infection' named entity 'time' named entity 'control' named entity 'systems' named entity 'E. coli' named entity 'strategy' named entity 'host' named entity 'efficient' named entity 'gRNA' named entity 'highly' named entity 'points' named entity 'strain' named entity 'strain' named entity 'created' named entity 'strains' named entity 'Escherichia coli' named entity 'E. coli' named entity 'Escherichia coli' named entity '2.0' named entity 'phage' named entity 'RFP' named entity 'phage' named entity 'phage' named entity 'CRISPR' named entity 'Cas9' named entity 'infection' named entity 'CRISPR' named entity 'phage' named entity 'CRISPR' named entity 'CRISPR/Cas9' named entity 'infection' named entity 'abortive infection' named entity 'loci' named entity 'safe and effective' named entity 'tryptone' named entity 'phages' named entity 'loci' named entity 'infection' named entity 'pBad' named entity 'Cas9' named entity 'vector' named entity 'phages' named entity 'life cycle' named entity 'phage' named entity 'CRISPR' named entity 'fluorescence' named entity 'plasmids' named entity 'E. coli' named entity 'phage' named entity 'CRISPR' named entity 'E. coli' named entity 'CRISPR' named entity 'fluorescence' named entity 'Fisher Scientific' named entity '3.8' named entity 'phage' named entity 'E. coli' named entity 'infection' named entity 'Cas9' named entity 'cas9' named entity 'heterologous proteins' named entity 'phage' named entity 'phage particles' named entity 'CRISPR' named entity 'Cas9'

Faceted Search & Find service v1.13.91 as of Mar 24 2020

Alternative Linked Data Documents: Sponger | ODE Content Formats:

RDF

ODATA

Microdata

About

OpenLink Virtuoso version 07.20.3229 as of Jul 10 2020, on Linux (x86_64-pc-linux-gnu), Single-Server Edition (94 GB total memory)
Data on this page belongs to its respective rights holders.
Virtuoso Faceted Browser Copyright © 2009-2024 OpenLink Software