The silencing of cysteine proteases in Fasciola hepatica newly excysted juveniles using RNA interference reduces gut penetration.
Probing protein function in parasitic flatworms is hampered by the difficulties associated with the development of transgenic approaches. Although RNA interference (RNAi) in schistosomes shows much promise, it has not been reported in other trematodes. Here, we show the successful silencing of the cysteine proteases cathepsin B and L in the infective stage of Fasciola hepatica newly excysted juveniles (NEJs). Silencing resulted in marked reductions in target transcript levels and significant diminution in the encoded proteins in the gut. RNAi of either enzyme in NEJs induced transient, abnormal locomotory phenotypes, and significantly reduced penetration of the rat intestinal wall.
Biomolecular Processes/Parasitology, School of Biological Sciences, Queen's University Belfast, Medical Biology Centre, 97 Lisburn Road, Belfast BT9 7BL, UK.
Louise
L
McGonigle
Angela
A
Mousley
Nikki J
NJ
Marks
Gerard P
GP
Brennan
John P
JP
Dalton
Terry W
TW
Spithill
Tim A
TA
Day
Aaron G
AG
Maule
D000818:Animals; D002401:Cathepsin B / Q000235:genetics; D056668:Cathepsin L; D002403:Cathepsins / Q000235:genetics; D003546:Cysteine Endopeptidases / Q000235:genetics*; D005210:Fasciola hepatica / Q000201:enzymology* / Q000502:physiology; D005211:Fascioliasis / Q000201:enzymology / Q000628:therapy*; D041981:Gastrointestinal Tract / Q000469:parasitology; D020868:Gene Silencing; D015316:Genetic Therapy / Q000379:methods*; D007411:Intestinal Diseases, Parasitic / Q000201:enzymology / Q000628:therapy*; D008124:Locomotion; D034622:RNA Interference; D051381:Rats; D017208:Rats, Wistar
D002403:Cathepsins; D003546:Cysteine Endopeptidases; D002401:Cathepsin B; D056668:Cathepsin L; C534280:Ctsl protein, rat