TY  - JOUR
TI  - Generation, Propagation, and Titering of Dicistrovirus From an Infectious Clone
AU  - Shen, Junzhou 
AU  - Sadasivan, Jibin 
AU  - Jan, Eric 
VL  - 15
IS  - 4
PY  - 2025
DA  - 2025/02/20
SP  - e5216
C1  - Bio-protocol 2025;15:e5216
DO  - 10.21769/BioProtoc.5216
UR  - https://doi.org/10.21769/BioProtoc.5216
AB  - Cricket paralysis virus (CrPV), a member of the family Dicistroviridae, is a single-stranded positive-sense RNA virus that primarily infects arthropods. Some members of the dicistrovirus family, including the honey bee viruses Israeli acute paralysis virus and Acute bee paralysis virus and the shrimp-infecting Taura syndrome virus, pose significant threats to agricultural ecosystems and economies worldwide. Dicistrovirus infection in Drosophila is used as a model system to study fundamental insect–virus–host interactions. The availability of a CrPV infectious clone allows controlled manipulation of the viral genome at a molecular level. Effective viral propagation and titration techniques are crucial for understanding the pathogenesis and epidemiology of dicistrovirus infections. Traditional methods for assessing viral titers, such as plaque assays, are unsuitable for CrPV, since Drosophila tissue culture cells like Schneider 2 cells cannot readily form adherent plaques. Here, we present a streamlined protocol for generating a recombinant virus from a CrPV infectious clone, propagating the virus in S2 cells and titering the virus by an immunofluorescence-based focus-forming assay (FFA). This protocol offers a rapid and reliable approach for generating recombinant viruses, viral amplification, and determining CrPV titers, enabling efficient investigation into viral biology and facilitating the development of antiviral strategies.
KW  - Cricket paralysis virus
KW  - Infectious clone
KW  - Amplification
KW  - Dicistrovirus
KW  - Infection
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False