Molecular characterization, tissue-specific expression and RNAi knockdown of the first vitellogenin receptor from a tick

Mitchell, Robert D.; Ross, Elizabeth; Osgood, C.; Sonenshine, Daniel E.; Donohue, Kevin V.; Khalil, Sayed M.S.; Thompson, Deborah; Roe, R. Michael

doi:10.1016/j.ibmb.2007.01.005

Cited by 65 publications

(47 citation statements)

References 32 publications

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“…In brief, this course taught by different instructors successfully met its learning objectives. Furthermore, after the completion of the course, one student successfully designed and executed an RNAi experiment in his own research on the American dog tick, Dermacentor variabilis (Mitchell et al, 2007), and has recently accepted a position in a biotechnology company to continue performing research using RNAi.…”

Section: Positive Course Outcomesmentioning

confidence: 99%

A Laboratory-intensive Course on RNA Interference and Model Organisms

Miller

Witherow

Carson

2009

LSE

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RNA interference (RNAi) is a powerful method to silence gene expression in a variety of organisms and is generating interest not only as a useful tool for research scientists but also as a novel class of therapeutics in clinical trials. Here, we report that undergraduate and graduate students with a basic molecular biology background were able to demonstrate conceptual knowledge and technical skills for using RNAi as a research tool upon completion of an intensive 8-wk RNAi course with a 2-h lecture and 5-h laboratory per week. Students were instructed on design of RNAi experiments in model organisms and perform multiweek laboratory sessions based on journal articles read and discussed in class. Using Nicotiana benthamiana, Caenorhabditis elegans, and mammalian cell culture, students analyzed the extent of silencing using both qualitative assessment of phenotypic variations and quantitative measurements of RNA levels or protein levels. We evaluated the course over two semesters, each with a separate instructor. In both semesters, we show students met expected learning outcomes as demonstrated by successful laboratory experiment results, as well as positive instructor assessments of exams and lab reports. Student self-assessments revealed increased confidence in conceptual knowledge and practical skills. Our data also suggest that the course is adaptable to different instructors with varying expertise.

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Section: Positive Course Outcomesmentioning

confidence: 99%

A Laboratory-intensive Course on RNA Interference and Model Organisms

Miller

Witherow

Carson

2009

LSE

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“…Silencing these genes through RNAi greatly reduced the reproductive capacity of female ticks, which showed immature and light-colored oocytes. The uptake of vitellogenin in the oocytes is facilitated by vitellogenin receptor, which has been characterized in D. variabilis [34], H.longicornis [99], and A. hebraeum [100]. Aside from the negative impact in oviposition consistently induced by RNAi in all these studies, silencing of H. longicornis vitellogenin receptor also reportedly inhibited the transovarial transmission of Babesia gibsoni.…”

Section: Genes Related To Reproductive Functionmentioning

confidence: 79%

“…Through this method, dsRNA is usually introduced directly into the hemocoel of ticks allowing the dsRNA to circulate within the hemolymph. In most reports, a high concentration of at least 1 µg dsRNA per tick has been shown to be effective in inducing gene silencing [33], but in some reports, lower concentration has been found to be similarly effective [34][35][36]. Injection has been accomplished using a 33-36-gauge needle attached to a Hamilton syringe particularly in large tick species, such as Amblyomma americanum [37], Dermacentor variabilis, and D. marginatus [38], while microinjection using a microcapillary drawn to a fine point needle and inserted to a micromanipulator has been commonly employed in smaller tick species, including Ixodes [39], and Haemaphysalis [26] ticks.…”

Section: Injectionmentioning

confidence: 99%

RNA Interference – A Powerful Functional Analysis Tool for Studying Tick Biology and its Control

Galay¹,

Umemiya‐Shirafuji²,

MasamiMochizuki³

et al. 2016

RNA Interference

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Ticks (Acari: Ixodida) are blood-sucking arthropods globally recognized as vectors of numerous diseases. They are primarily responsible for the transmission of various pathogens, including viruses, rickettsiae, and blood parasites of animals. Ticks are second to mosquitoes in terms of disease transmission to humans. The continuous emergence of tick-borne diseases and acaricide resistance of ticks necessitates the development of new and more effective control agents and strategies; therefore, understanding of different aspects of tick biology and their interaction with pathogens is very crucial in developing effective control strategies. RNA interference (RNAi) has been widely used in the area of tick research as a versatile reverse genetic tool to elucidate the functions of various tick proteins. During the past decade, numerous studies on ticks utilized RNAi to evaluate potentially key tick proteins involved in blood feeding, reproduction, evasion of host immune response, interaction with pathogens, and pathogen transmission that may be targeted for tick and pathogen control. This chapter reviewed the application of RNAi in tick research over the past decade, focusing on the impact of this technique in the advancement of knowledge on tick and pathogen biology.

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“…After synthesis, Vg is released into haemolymph and taken up via receptor-mediated endocytosis by the growing oocytes. Vg is partly processed in the endosomal compartment and then stored as Vt in specialized organelles called yolk granules (Raikhel and Dhadialla, 1992;Fagotto, 1995;Mitchell et al 2007).…”

Section: Introductionmentioning

confidence: 99%

Localization and function ofRhipicephalus (Boophilus) microplusvitellin-degrading cysteine endopeptidase

et al. 2010

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S U M M A R YThe tick Rhipicephalus (Boophilus) microplus is an important parasite of cattle in many areas of the tropics. Characterization of molecules involved in mechanisms such as vitellogenesis and embryo development may contribute to a better understanding of this parasite's physiology. The vitellin-degrading cysteine endopeptidase (VTDCE) is the most active enzyme involved in vitellin hydrolysis in R. microplus eggs. Here we show an association between VTDCE and vitellin in an additional site, apart from the active site. Our data also demonstrate cysteine endopeptidase activity in different tissues such as ovary, gut, fat body, salivary gland and female haemolymph, where it is controlled by a physiological inhibitor. In R. microplus female gut, VTDCE is localized in areas of protein synthesis and trafficking with the underlying haemolymph. VTDCE is also localized in the ovary basal region, in vesicle membranes of ovary pedicel cells and in oocyte cytosol. These results suggest that VTDCE plays a role in vitellin digestion during tick development.

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Molecular characterization, tissue-specific expression and RNAi knockdown of the first vitellogenin receptor from a tick

Cited by 65 publications

References 32 publications

A Laboratory-intensive Course on RNA Interference and Model Organisms

A Laboratory-intensive Course on RNA Interference and Model Organisms

RNA Interference – A Powerful Functional Analysis Tool for Studying Tick Biology and its Control

Localization and function ofRhipicephalus (Boophilus) microplusvitellin-degrading cysteine endopeptidase

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