Prof.Zhou Hongbo from College of Animal Sciences & Technology of HZAU will get direct funds of 2.55 million yuan from Sino-British NSFC-RCUK_BBSRC Program, with his project “The Identification and Function of Host Genes Required for the Replication of Swine Influenza Virus” in the field of Animal Health. It is a cooperative research project between National Natural Science Foundation of China (NSFC)and BBSRC, UK’s Biotechnology and Biological Sciences Research Council (NSTDA), Philippine Department of Agriculture Fisheries Biotechnology Project (DA-BIOTECH), Philippine Department of Bureau of Agricultural Research (DA-BAR).
Swine flu is one of the most prevalent and serious diseases in large-scale pig farms, which poses a serious threat to pig breading and public health. Recently, through gene editing techniques, scientists have successfully generated the first gene-edited pig that is resistant to infection with the swine pathogen PRRS. Based on this, HZAU will collaborate with Imperial College London in the project to discover and characterize the swine host factors required to support swine influenza virus replication. The result will lay the foundation for developing the gene edited pig resistant to SIV. First, the researchers will develop a pig Genome-scale CRISPR knockout (GeCKO) library. Then, the lentiviral sgRNA library will be transduced into the pre-constructed Cas9 expressing newborn pig trachea (NPTr) cells and a survival screening strategy will be carried out to identify genes essential for SIV-induced killing. In addition to the survival screen we will also develop a screen for viral HA expression levels that will aim to identify genes required for replication rather than those for virus entry. The sgRNAs in the surviving or sorting cells will be PCR-amplified and deep sequenced using Illumina second-generation sequencing to identify the genos that had been targeted. To map domains and amino acids of the genes that are required for their support of SIV replication, the researchers will introduce the wild type or mutant versions of each protein by plasmid transfection back into the CRISPR cells to complement polymerase activity. In addition researchers will develop and employ a high throughput condon library mutagenesis screen, combined with FACS sourcing and deep sequencing, to probe the entire evolutionary space of the host protein for this function. The gene editing NPTr with a tagged mutant protein will be developed. It also used the CRISPR technology to produce tagged cell lines with mutation in key loca of essential genes. These cell lines were used to identified mutant’s influence on virus replication and to further clarify their molecular mechanism. The project is not only helpful to further clarify the molecular mechanism of the influenza virus transcription and replication but also can spot antiviral drug targets from host system, thus providing new ideas for the research and development of anti-influenza drugs.
Original Text: http://news.hzau.edu.cn/2017/1229/51194.shtml
Tr. by Lin Zhenhua;
Instructor: Guo Haiyan
