Capillary feeding of specific dsRNA induces silencing of the isac gene in nymphal Ixodes scapularis ticks.
Ixodes scapularis transmits several pathogens including Borrelia burgdorferi. Bioactive compounds in tick saliva support tick feeding and influence pathogen transmission to the mammalian host. These studies utilized oral delivery of dsRNA to silence an anticomplement gene (isac) in I. scapularis nymphs. Silencing of isac significantly reduced fed-tick weight compared to delivery of control lacZ dsRNA, and immunoblots specific for FlaB protein indicated a reduction in spirochete load in isac-silenced infected nymphs. SDS-PAGE demonstrated that isac gene silencing affected expression of a number of salivary and non-salivary gland proteins in ticks. Finally, multiple isac cDNA homologues were cloned, and these may represent a new gene family coexpressed during tick feeding. This work presents a novel oral delivery approach for specific gene silencing in I. scapularis nymphs and characterizes the effect of isac on blood-feeding in an attempt to block transmission of B. burgdorferi.
Soares
C A G
CA
Centers for Disease Control and Prevention, Division of Vector-Borne Infectious Diseases, Bacterial Zoonoses Branch, Foothills Campus, Fort Collins, CO 80522, USA.
Lima
C M R
CM
Dolan
M C
MC
Piesman
J
J
Beard
C B
CB
Zeidner
N S
NS
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D025065:Borrelia burgdorferi / Q000254:growth & development; D003001:Cloning, Molecular; D020868:Gene Silencing; D018884:Ixodes / Q000235:genetics* / Q000382:microbiology; D008193:Lyme Disease / Q000635:transmission; D051379:Mice; D008969:Molecular Sequence Data; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000235:genetics* / Q000502:physiology; D034741:RNA, Small Interfering / Q000235:genetics / Q000502:physiology; D012471:Salivary Proteins and Peptides / Q000096:biosynthesis / Q000235:genetics*; D016415:Sequence Alignment; D013047:Specific Pathogen-Free Organisms
C411881:Isac protein, Ixodes scapularis; D012330:RNA, Double-Stranded; D034741:RNA, Small Interfering; D012471:Salivary Proteins and Peptides