Retroviral delivery of RNA interference against Marek's disease virus in vivo.
The process of RNA interference (RNAi) has been exploited in cultured chicken cells and in chick embryos to assess the effect of specific gene inhibition on phenotypes related to development and disease. We previously demonstrated that avian leukosis virus-based retroviral vectors are capable of delivering effective RNAi against Marek's disease virus (MDV) in cell culture. In this study, similar RNAi vectors are shown to reduce the replication of MDV in live chickens. Retroviral vectors were introduced into d 0 chick embryos, followed by incubation until hatching. Chicks were challenged with 500 pfu of strain 648A MDV at day of hatch, followed by assays for viremia at 14 d postinfection. Birds were monitored for signs of Marek's disease for 8 wk. A stem-loop PCR assay was developed to measure siRNA expression levels in birds. Delivery of RNAi co-targeting the MDV gB glycoprotein gene and ICP4 transcriptional regulatory gene significantly reduced MDV viremia in vivo, although to lesser extents than were observed in cell culture. Concomitant reductions in disease incidence also were observed, and the extent of this effect depended on the potency of the MDV challenge virus inoculum. Successful modification of phenotypic traits in live birds with retroviral RNAi vectors opens up the possibility that such approaches could be used to alter the expression of candidate genes hypothesized to influence a variety of quantitative traits including disease susceptibility.
Chen
M
M
Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing 48824, USA.
Payne
W S
WS
Dunn
J R
JR
Chang
S
S
Zhang
H M
HM
Hunt
H D
HD
Dodgson
J B
JB
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002642:Chick Embryo; D002645:Chickens; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D025922:Mardivirus / Q000502:physiology*; D034622:RNA Interference; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000378:metabolism; D014779:Virus Replication
D014764:Viral Proteins