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CRISPR screening of porcine sgRNA library identifies host factors associated with Japanese encephalitis virus replication

ABSTRACT: CRISPR screening of porcine sgRNA library identifies host factors associated with Japanese encephalitis virus replication

PROVIDER: PRJNA664556 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR12677926_1.fastq.gz	Fastqsanger.gz
	SRR12677926_2.fastq.gz	Fastqsanger.gz
	SRR12677927_1.fastq.gz	Fastqsanger.gz
	SRR12677927_2.fastq.gz	Fastqsanger.gz
	SRR12677928_1.fastq.gz	Fastqsanger.gz

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CRISPR screening of porcine sgRNA library identifies host factors associated with Japanese encephalitis virus replication

Project description:We design a porcine genome-scale CRISPR/Cas9 knockout (PigGeCKO) library containing 85,674 single guide RNAs targeting 17,743 protein-coding genes, 11,053 long ncRNAs, and 551 microRNAs. Subsequently, we use the PigGeCKO library to identify key host factors facilitating JEV infection in porcine cells.

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CRISPR screening of porcine sgRNA library identifies host factors associated with Japanese encephalitis virus replication.

Project description:Japanese encephalitis virus (JEV) is a mosquito-borne zoonotic flavivirus that causes encephalitis and reproductive disorders in mammalian species. However, the host factors critical for its entry, replication, and assembly are poorly understood. Here, we design a porcine genome-scale CRISPR/Cas9 knockout (PigGeCKO) library containing 85,674 single guide RNAs targeting 17,743 protein-coding genes, 11,053 long ncRNAs, and 551 microRNAs. Subsequently, we use the PigGeCKO library to identify key host factors facilitating JEV infection in porcine cells. Several previously unreported genes required for JEV infection are highly enriched post-JEV selection. We conduct follow-up studies to verify the dependency of JEV on these genes, and identify functional contributions for six of the many candidate JEV-related host genes, including EMC3 and CALR. Additionally, we identify that four genes associated with heparan sulfate proteoglycans (HSPGs) metabolism, specifically those responsible for HSPGs sulfurylation, facilitate JEV entry into porcine cells. Thus, beyond our development of the largest CRISPR-based functional genomic screening platform for pig research to date, this study identifies multiple potentially vulnerable targets for the development of medical and breeding technologies to treat and prevent diseases caused by JEV.

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Project description:Japanese encephalitis (JE) is an acute encephalitis syndrome contributed to Japanese encephalitis virus (JEV) infection. It is the chief cause of viral encephalitis in Asian. In recent years, association of JEV infection with neurological problems such as Guillain-Barré syndrome had reported. Nevertheless, its potential pathogenic mechanism has never been reported. Therefore, it is urgent to explore the relationship between peripheral nerve injury and JEV infection. Here, we use the liquid chromatography-tandem mass spectrometry (LC-MS/MS) technique to make out the protein expression levels of mice sciatic nerve between JEV infection group and the sham group. In general, 4303 proteins were identified by MS, and 187 differentially expressed proteins were found. There were 105 proteins up-regulated in the injured sciatic nerve, and 82 proteins were down-regulated. Functional enrichment analysis of differentially expressed proteins showed that the up-regulated proteins were mainly related to immune regulatory response, and the down-regulated proteins were related to ribosomal structural components and translation.

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