CRISPR/Cas9-mediated multiple single guide RNAs potently abrogate pseudorabies virus replication

被引:32
|
作者
Tang, Yan-Dong [1 ]
Liu, Ji-Ting [1 ]
Wang, Tong-Yun [1 ]
Sun, Ming-Xia [1 ]
Tian, Zhi-Jun [1 ]
Cai, Xue-Hui [1 ]
机构
[1] Chinese Acad Agr Sci, State Key Lab Vet Biotechnol, Harbin Vet Res Inst, 678 HaPing Rd, Harbin 150069, Heilongjiang, Peoples R China
关键词
CRISPR/Cas9; Pseudorabies virus; Inhibition; SgRNA; INHIBIT HIV-1 REPLICATION; GENOMIC CHARACTERIZATION; CELLS; CHINA; GENE; ENDONUCLEASE; VARIANT; ESCAPE; INFECTION; STRATEGY;
D O I
10.1007/s00705-017-3553-4
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Pseudorabies virus (PRV) is a swine herpesvirus that causes significant morbidity and mortality in swine populations and has caused huge economic losses in the worldwide swine industry. Currently, there is no effective antiviral drug in clinical use for PRV infection; it is also difficult to eliminate PRV from infected swine. In our study, we set out to combat these swine herpesvirus infections by exploiting the CRISPR/Cas9 system. We designed 75 single guide RNAs (sgRNA) by targeting both essential and non-essential genes across the genome of PRV. We applied a firefly luciferase-tagged reporter PRV virus for high-throughput sgRNA screening and found that most of the sgRNAs significantly inhibited PRV replication. More importantly, using a transfection assay, we demonstrated that simultaneous targeting of PRV with multiple sgRNAs completely abolished the production of infectious viruses in cells. These data suggest that CRISPR/Cas9 could be a novel therapeutic agent against PRV in the future.
引用
收藏
页码:3881 / 3886
页数:6
相关论文
共 50 条
  • [41] CRISPR–Cas9-mediated chromosome engineering in Arabidopsis thaliana
    Michelle Rönspies
    Patrick Schindele
    Rebecca Wetzel
    Holger Puchta
    Nature Protocols, 2022, 17 : 1332 - 1358
  • [42] CRISPR/Cas9-mediated ssDNA Recombineering in Corynebacterium glutamicum
    Liu, Jiao
    Wang, Yu
    Zheng, Ping
    Sun, Jibin
    BIO-PROTOCOL, 2018, 8 (19):
  • [43] CRISPR/Cas9-mediated Immunity in Plants Against Pathogens
    Sameeullah, Muhammad
    Khan, Faheem Ahmed
    Ozer, Goksel
    Aslam, Noreen
    Gurel, Ekrem
    Waheed, Mohammad Tahir
    Karadeniz, Turan
    CURRENT ISSUES IN MOLECULAR BIOLOGY, 2018, 26 : 55 - 63
  • [44] CRISPR/Cas9-Mediated Mutagenesis of RCO in Cardamine hirsuta
    Alvim Kamei, Claire Lessa
    Pieper, Bjorn
    Laurent, Stefan
    Tsiantis, Miltos
    Huijser, Peter
    PLANTS-BASEL, 2020, 9 (02):
  • [45] CRISPR/Cas9-mediated targeted mutagenesis in Nicotiana tabacum
    Junping Gao
    Genhong Wang
    Sanyuan Ma
    Xiaodong Xie
    Xiangwei Wu
    Xingtan Zhang
    Yuqian Wu
    Ping Zhao
    Qingyou Xia
    Plant Molecular Biology, 2015, 87 : 99 - 110
  • [46] CRISPR/Cas9-mediated targeted mutagenesis in Nicotiana tabacum
    Gao, Junping
    Wang, Genhong
    Ma, Sanyuan
    Xie, Xiaodong
    Wu, Xiangwei
    Zhang, Xingtan
    Wu, Yuqian
    Zhao, Ping
    Xia, Qingyou
    PLANT MOLECULAR BIOLOGY, 2015, 87 (1-2) : 99 - 110
  • [47] CRISPR/Cas9-mediated correction of human genetic disease
    Ke Men
    Xingmei Duan
    Zhiyao He
    Yang Yang
    Shaohua Yao
    Yuquan Wei
    Science China(Life Sciences) , 2017, (05) : 447 - 457
  • [48] CRISPR/Cas9-Mediated Mutagenesis of Antennapedia in Spodoptera frugiperda
    Wang, Congke
    Zhao, Te
    Liu, Xiaolong
    Li, Tianliang
    He, Leiming
    Wang, Qinqin
    Wang, Li
    Zhou, Lin
    INSECTS, 2024, 15 (01)
  • [49] Genetic chimerism of CRISPR/Cas9-mediated rice mutants
    Jang, Geupil
    Lee, Sangyool
    Um, Tae Young
    Chang, Sun Hyun
    Lee, Han Yong
    Chung, Pil Joong
    Kim, Ju-Kon
    Do Choi, Yang
    PLANT BIOTECHNOLOGY REPORTS, 2016, 10 (06) : 425 - 435
  • [50] CRISPR/Cas9-mediated correction of human genetic disease
    Ke Men
    Xingmei Duan
    Zhiyao He
    Yang Yang
    Shaohua Yao
    Yuquan Wei
    Science China Life Sciences, 2017, 60 : 447 - 457