Chengliang Gong

Soochow University

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Guangli Cao

Soochow University

113

Renyu Xue

Linyi People's Hospital

113

Xiaolong Hu

Jiangxi Agricultural University

Min Zhu

Nanchang University

Zi Liang

Soochow University

Jun Pan

Agricultural Genomics Institute at Shenzhen

Dhiraj Kumar

International Centre for Genetic Engineering and Biotechnology

Yongjie Feng

Soochow University

Liyuan Zhu

Chinese Academy of Medical Sciences & Peking Union Medical College

Xinyu Tong

Soochow University

Cooperative Institutions

Soochow University

137

Chinese Academy of Sciences

Sun Yat-sen University

Shanghai Jiao Tong University

Zhejiang University

Huazhong University of Science and Technology

Chinese Academy of Medical Sciences & Peking Union Medical College

Peking University

Ministry of Agriculture and Rural Affairs

Capital Medical University

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CRISPR/Cas9 Initiated Transgenic Silkworms as a Natural Spinner of Spider Silk

Biomacromolecules (2019)

Xiaoli Zhang Lijin Xia Breton A. Day Thomas I. Harris Paula F. Oliveira

Using transgenic silkworms with their natural spinning apparatus has proven to be a promising way to spin spider silk-like fibers. The challenges are incorporating native-size spider silk proteins and achieving an inheritable transgenic silkworm strain. In this study, a CRISPR/Cas9 initiated fixed-point strategy was used to successfully incorporate spider silk protein genes into the Bombyx mori genome. Native-size spider silk genes (up to 10 kb) were inserted into an intron of the fibroin heavy or light chain (FibH or FibL) ensuring that any sequence changes induced by the CRISPR/Cas9 would not impact protein production. The resulting fibers are as strong as native spider silks (1.2 GPa tensile strength). The transgenic silkworms have been tracked for several generations with normal inheritance of the transgenes. This strategy demonstrates the feasibility of using silkworms as a natural spider silk spinner for industrial production of high-performance fibers.

Spider Silk

Fibroin

10.1021/acs.biomac.9b00193

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Citations (61)

Evaluation of immune efficacy of GCRV vp6 DNA vaccine

Zhongguo Shuichan Kexue (2012)

Lin Liu XU Shi-ying Jinghui Li Yong Zou Jindi Ni

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Construction and immune efficacy studies on DNA vaccine with VP7 gene of grass carp reovirus

Xu Shiying Lin Liu Jinghui Li Yong Zou Jindi Ni

Double GCRV outer capsid protein VP7 genes(0.9 kb)and β-actin promoter(0.56 kb)from Megalobrama amblycephala were amplified and cloned into gene transfer vector pFastBacTM Dual to obtain recombinant pFastBac-β-VP71-VP72.Immune experiment and immunological effect were studied.The recombinant plasmids were divided into three groups based on doses:10,30,60 μg.30 μg pFastBacTM Dual group and control group were designed in addition.Immune efficacy was determined by means of transcriptions of VP7,antibody detection by indirect agglutination reaction and challenge experiment on the 14th,21st,28th,and 49th days after immunization.The results showed that pFastBac-β-VP71-VP72 was transformed into body cell,VP7 gene could be controlled for expression constantly by β-actin promoter,and transcription of VP7 was still in process on the 49th day.Antibody was produced and the titers reached their peak on the 21th day after immunization.The mortality rates were 0%,0% and 5% respectively in 10 μg,30 μg and 60 μg vaccine immunization groups,30% in pFastBacTM Dual group,and 100% in control group after GCRV injection.All results suggested that pFastBac-β-VP71-VP72 had good immuno-protective efficacy as DNA vaccine against GCRV.Therefore,the study provided important academic basis for research,development and application of the GCRV genetic vaccine.

Antibody titer

Grass carp

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BmCPV replication is suppressed by the activation of the NF-κB/autophagy pathway through the interaction of vsp21 translated by vcircRNA_000048 with ubiquitin carboxyl-terminal hydrolase

Insect Biochemistry and Molecular Biology (2023)

Yunshan Zhang Min Zhu Jun Pan Qunnan Qiu Xinyu Tong

Deubiquitinating enzyme

Viral protein

10.1016/j.ibmb.2023.103947

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Citations (3)

Baculoviruses as A Novel Gene-delivery Vector for Mammalian Cell

Chengliang Gong

The baculoviruses have been used for expression of many kinds of recombinant proteins in insect cell by using the polyhedrin gene promoter. Recently, many commonly used cell lines as well as primary cultures are efficiently transduced with baculovirus, and little cytotoxicity could be observed. Recombinant baculoviruses vectors engineered to contain mammalian cell active promoter elements, have been used successfully for transient and stable gene delivery in mammalian cells, baculoviruses as gene therapy xenovectors, are able to deliver target gene to mammalian cells, and induce special immunoreactions directionally to expression product of the gene. The efficiency to deliver gene to mammalian cell with baculoviruses was inhibited obviously by complement cmponents, while pseudotyped baculoviruses display complement resistance. Gene delivery mediated by baculovirus is a powerful tool to estimate the strength of promoter in different cell. Hybrid baculoviruses containing the genomes of heterologous viruses were able to launch viral infections; this method has been used to examine effects of anti viral drugs. Baculoviruses as safe, efficient, and specific gene delivery vectors hold great promise for the development of a new generation of more efficient gene therapy vehicles in the future.

Polyhedrin

Heterologous

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Viral Circular RNAs and Their Possible Roles in Virus-Host Interaction

Frontiers in Immunology (2022)

Xing Zhang Zi Liang Chonglong Wang Zeen Shen Sufei Sun

Circular RNAs (circRNAs) as novel regulatory molecules have been recognized in diverse species, including viruses. The virus-derived circRNAs play various roles in the host biological process and the life cycle of the viruses. This review summarized the circRNAs from the DNA and RNA viruses and discussed the biogenesis of viral and host circRNAs, the potential roles of viral circRNAs, and their future perspective. This review will elaborate on new insights gained on viruses encoded circRNAs during virus infection.

Viral life cycle

DNA virus

Circular RNA

Host factors

10.3389/fimmu.2022.939768

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Citations (20)

Grass carp reovirus encoding circular RNAs with antiviral activity

Aquaculture (2020)

Jun Pan Xing Zhang Yunshan Zhang Bingyu Yan Kun Dai

Grass carp

RNA Silencing

Circular RNA

MDA5

10.1016/j.aquaculture.2020.736135

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Citations (17)

Tight junction protein claudin-2 promotes cell entry of Bombyx mori cypovirus

Applied Microbiology and Biotechnology (2021)

Min Zhu Xing Zhang Jun Pan Hanxue Zhu Ziyao Zhang

Claudin

10.1007/s00253-021-11456-y

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Citations (4)

Metatranscriptomic Analysis Reveals an Imbalance of Hepatopancreatic Flora of Chinese Mitten Crab Eriocheir sinensis with Hepatopancreatic Necrosis Disease

Biology (2021)

Zeen Shen Dhiraj Kumar Xunmeng Liu Bingyu Yan Ping Fang

Hepatopancreas necrosis disease (HPND) of the Chinese mitten crab Eriocheir sinensis causes huge economic loss in China. However, the pathogenic factors and pathogenesis are still a matter of dissension. To search for potential pathogens, the hepatopancreatic flora of diseased crabs with mild symptoms, diseased crabs with severe symptoms, and crabs without visible symptoms were investigated using metatranscriptomics sequencing. The prevalence of Absidia glauca and Candidatus Synechococcus spongiarum decreased, whereas the prevalence of Spiroplasma eriocheiris increased in the hepatopancreatic flora of crabs with HPND. Homologous sequences of 34 viral species and 4 Microsporidian species were found in the crab hepatopancreas without any significant differences between crabs with and without HPND. Moreover, DEGs in the hepatopancreatic flora between crabs with severe symptoms and without visible symptoms were enriched in the ribosome, retinol metabolism, metabolism of xenobiotics by cytochrome P450, drug metabolism—cytochrome P450, biosynthesis of unsaturated fatty acids, and other glycan degradation. Moreover, the relative abundance of functions of DEDs in the hepatopancreatic flora changed with the pathogenesis process. These results suggested that imbalance of hepatopancreatic flora was associated with crab HPND. The identified DEGs were perhaps involved in the pathological mechanism of HPND; nonetheless, HPND did not occur due to virus or microsporidia infection.

Hepatopancreas

Chinese mitten crab

10.3390/biology10060462

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Citations (15)

Identification and characterization of a novel small viral peptide (VSP59) encoded by Bombyx mori cypovirus (BmCPV) that negatively regulates viral replication

Microbiology Spectrum (2024)

Manman Cao Qunnan Qiu Xing Zhang Wenxue Zhang Zeen Shen

ABSTRACT Bombyx mori cypovirus (BmCPV), a member of the Reoviridae family, is a well-established research model for double-stranded RNA (dsRNA) viruses with segmented genomes. Despite its small genome size, the coding potential of BmCPV remains largely unexplored. In this study, we identified a novel small open reading frame within the S10 dsRNA genome, encoding a small viral peptide (VSP59) with 59 amino acid residues. Functional characterization revealed that VSP59 acts as a negative regulator of viral replication. VSP59 predominantly localizes to the cytoplasm, where it interacts with prohibitin 2 (PHB2), an inner membrane mitophagy receptor. This interaction targets mitochondria and triggers caspase 3-dependent apoptosis. Transient expression of vsp59 in BmN cells suppressed viral replication, an effect that was reversed by silencing PHB2 expression. Moreover, recombinant BmCPV with a mutated vsp59 exhibited reduced replication. Our findings demonstrate that VSP59 interacts with PHB2 on mitochondria, inducing apoptosis and thereby diminishing viral replication. This study expands our understanding of the genetic information encoded by the BmCPV genome and highlights the role of novel small peptides in host-virus interactions. IMPORTANCE A novel small open reading frame (sORF) from the viral genome was identified and characterized. The sORF could encode a small viral peptide (VSP59) that targeted mitochondria and induced prohibitin 2-related apoptosis, further attenuating Bombyx mori cypovirus replication.

RNA Silencing

10.1128/spectrum.00826-24

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Citations (1)