Autocidal control of ticks by silencing of a single gene by RNA interference

doi:10.1016/j.bbrc.2006.03.109

Biochemical and Biophysical Research Communications

Volume 344, Issue 1, 26 May 2006, Pages 332-338

https://doi.org/10.1016/j.bbrc.2006.03.109 Get rights and content

Abstract

Ticks impact human and animal health worldwide and new control methods are needed to circumvent drawbacks of tick control by acaricide application including selection of drug resistant ticks and environmental pollution. Using RNA interference we silenced the expression of a single gene, subolesin, and produced ticks with diminished reproductive performance and prevented successful mating and production of viable offspring. We propose a sterile acarine technique (SAT) for reduction of tick populations by release of subolesin-silenced ticks. Conservation of subolesin among tick species suggests that SAT may be useful for control of many medically and economically important tick species.

Section snippets

Materials and methods

Ticks. Adult ticks were obtained from the laboratory colony maintained at the Oklahoma State University Tick Rearing Facility. Sheep were housed at the Tick Rearing Laboratory with the approval and supervision of the OSU Institutional Animal Care and Use Committee. Off-host ticks were maintained in a 12 h light:12 h dark photoperiod at 22–25 °C and 95% relative humidity.

Generation of dsRNA. Oligonucleotide primers containing T7 promoter sequences for in vitro transcription and synthesis of dsRNA

Results and discussion

In an effort to devise new methods for tick control, we investigated the possibility of using RNAi to develop a SAT method for ticks, similar to the SIT commonly used successfully to suppress populations of economically important insect pests. Release of hybrid sterile ticks to mate with wild females that then produce unfit hybrid offspring was advanced as a putative alternative control method explored for the one-host Boophilus spp [8], [17], [18]. Although hybrid sterility can be considered a

Acknowledgments

The authors thank Drs. D. Sonenshine and S.A. Ewing for critical review of the manuscript. This research was supported by the Oklahoma Agricultural Experiment Station (project 1669), the Sitlington Endowed Chair for Food Animal Research (K.M.K). Consuelo Almazán was funded by Pfizer Animal Health, Kalamazoo, MI, and a Grant-in-Aid from the CONACYT and Promep (University of Tamaulipas), Mexico. V. Naranjo was funded by Consejería de Educación, JCCM, Spain.

References (24)

P. Parola et al.
Tick-borne bacterial diseases emerging in Europe
Clin. Microbiol. Infect.
(2001)
W.N. Beesley
The ecological basis of parasite control: ticks and flies
Vet. Parasitol.
(1982)
C. Almazán et al.
Identification of protective antigens for the control of Ixodes scapularis infestations using cDNA expression library immunization
Vaccine
(2003)
C. Almazán et al.
Characterization of three Ixodes scapularis cDNAs protective against tick infestations
Vaccine
(2005)
C. Almazán et al.
Vaccination with recombinant tick antigens for the control of Ixodes scapularis adult infestations
Vaccine
(2005)
S.C. Barker et al.
Systematics and evolution of ticks with a list of valid genus and species names
Parasitology
(2004)
Y.S. Balashov
Nutrition and course of spermatogenesis in male ixodids
Dokl. Akad. Nauk. USSR
(1956)
A. Estrada-Peña et al.
Ticks feeding on humans: a review of records on human-biting Ixodoidea with special reference to pathogen transmission
Exp. Appl. Acarol.
(1999)
K. Kocan et al.
Anaplasma marginale (Rickettsiales: Anaplasmataceae): recent advances in defining host-pathogen adaptations of a tick-borne rickettsia
Parasitology
(2004)
J. de la Fuente et al.
Advances in the identification and characterization of protective antigens for development of recombinant vaccines against tick infestations
Expert Rev. Vaccines
(2003)

P. Willadsen

Anti-tick vaccines

Parasitology

(2004)

F. Gould et al.

Population genetics of autocidal control and strain replacement

Annu. Rev. Entomol.

(2004)

Cited by (58)

Innovative approaches for the control of ticks and tick-borne diseases
2023, Ticks and Tick-borne Diseases
Ticks and tick-borne diseases constitute a major threat for human and animal health worldwide. Vaccines for the control of tick infestations and transmitted pathogens still represents a challenge for science and health. Vaccines have evolved with antigens derived from inactivated pathogens to recombinant proteins and vaccinomics approaches. Recently, vaccines for the control of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have shown the efficacy of new antigen delivery platforms. However, until now only two vaccines based on recombinant Bm86/Bm95 antigens have been registered and commercialized for the control of cattle-tick infestations. Nevertheless, recently new technologies and approaches are under consideration for vaccine development for the control of ticks and tick-borne pathogens. Genetic manipulation of tick commensal bacteria converted enemies into friends. Frankenbacteriosis was used to control tick pathogen infection. Based on these results, the way forward is to develop new paratransgenic interventions and vaccine delivery platforms for the control of tick-borne diseases.
Control of tick infestations in wild roe deer (Capreolus capreolus) vaccinated with the Q38 Subolesin/Akirin chimera
2020, Vaccine
Citation Excerpt :
The recombinant SUB/AKR chimera Q38 was amplified from synthetic genes (JX193856) optimized for codon usage in Escherichia coli and cloned into the expression vector pET101 and expressed in E. coli strain BL21 using the Champion pET101 Directional TOPO Expression kit (Carlsbad, CA, USA). Recombinant proteins produced using this expression system were fused to Histidine tags for purification by affinity to Ni [22,24]. Transformed E. coli strains were induced with IPTG for 4.5 h to produce recombinant chimeric Q38 protein, which were purified to > 85% of total cell proteins by Ni affinity chromatography using 1 ml HisTrap FF columns mounted on an AKTA–FPLC system (GE Healthcare, Piscataway, NJ, USA) in the presence of 7 M urea lysis buffer.
Ticks (Acari: Ixodidae) are considered to be the most important vectors of disease-causing pathogens in domestic and wild animals, and emerging and re-emerging tick-borne diseases (TBD) exert an enormous impact on them. Wild ungulates are hosts for a wide variety of tick species and tick-borne pathogens that affect human and animal health. Consequently, the control of tick infestations and tick-borne pathogen prevalence is essential in some regions. Acaricides and animal management or culling have been used for the control of tick infestations and TBD, but tick vaccines constitute the best alternative to reduce the impact of acaricides on tick resistance and the environment. Previous results of controlled vaccination trials have shown that the Q38 Subolesin/Akirin chimera containing conserved protective epitopes could be a candidate universal antigen to control multiple tick species infestations. Thus, vaccination trials are necessary to validate these results under field conditions. In this study, we characterized the effect of Q38 vaccine on a wild population of European roe deer (Capreolus capreolus) in the Andalusian roe deer Reference Station (Junta de Andalucía, Cádiz, Spain). In this location, roe deer suffer especially severe parasitic conditions in some periods and commercial pesticides and ixodicides that are authorized to control ticks without specificity are frequently applied in the field, posing a threat to the environment. Animals vaccinated over a three-year period showed an antibody response to the vaccine antigen and a reduction in tick infestations by multiple species including Hyalomma marginatum, H. lusitanicum, Rhipicephalus bursa and Ixodes ricinus previously identified in roe deer, when compared to untreated controls. These results suggest the efficacy of Q38 for the control of tick infestations in wildlife.
Controlling ticks and tick-borne diseases…looking forward
2018, Ticks and Tick-borne Diseases
Tick-borne diseases (TBDs) represent a growing burden for human and animal health worldwide. Several approaches including the use of chemicals with repellency and parasiticidal activity, habitat management, genetic selection of hosts with higher resistance to ticks, and vaccines have been implemented for reducing the risk of TBDs. However, the application of latest gene editing technologies in combination with vaccines likely combining tick and pathogen derived antigens and other control measures should result in the development of effective, safe, and environmentally sound integrated control programs for the prevention and control of TBDs. This paper is not a review of current approaches for the control of ticks and TBDs, but an opinion about future directions in this area.
Liposome mediated double-stranded RNA delivery to silence ribosomal protein P0 in the tick Rhipicephalus haemaphysaloides
2018, Ticks and Tick-borne Diseases
Control of ticks has been achieved primarily by the application of acaricides, which has drawbacks such as environmental contamination leading to the selection of pesticide-resistant ticks. The potential of dsRNA to suppress genes critical for tick survival due to its sequence specificity suggests that dsRNAs could be developed as tailor-made pesticides. In this study, the dsRNA of P0 gene from the tick, Rhipicephalus haemaphysaloides, was evaluated as a potential anti-tick agent. Effects of using different dsRNA delivery methods were tested by quantitative RT-PCR and tick bioassays to determine survival, feeding and reproduction. The results showed that P0 dsRNAs could be effectively delivered into ticks and silenced by incubating with liposomes. Incubation time was found to be the most important factor in dsRNA delivery and gene silencing compared with liposome types and dsRNA concentration. The effects of P0 dsRNA treatment on ticks were found to be significant on blood feeding, molting or reproduction. These data show that anti-tick agents based on dsRNAs could have potential use in tick control.
Targeting a global health problem: Vaccine design and challenges for the control of tick-borne diseases
2017, Vaccine
It has been over twenty years since the first vaccines for the control of tick infestations became commercially available. These vaccines proved their efficacy and the potential of this approach for the control of tick-borne diseases (TBDs), which represent a growing burden for human and animal health worldwide. In all these years, research in this area has produced new tick-derived and pathogen-derived candidate protective antigens. However, the potential of vaccines for the control of TBDs has been underestimated due to major challenges to reduce tick infestations, pathogen infection, multiplication and transmission, tick attachment and feeding time and/or host pathogen infection. Nevertheless, vaccines constitute the most safe and effective intervention for the control of TBDs in humans, domestic and wild animals.
DsRNA-mediated targeting of ribosomal transcripts RPS6 and RPL26 induces long-lasting and significant reductions in fecundity of the vector Aedes aegypti
2016, Journal of Insect Physiology
Ribosomal transcripts produce critical proteins that are involved in most cellular production processes. Targeting ribosomal transcripts has produced mortality in mites and ticks but the effect of ribosomal transcript knockdown has not been thoroughly examined in mosquitoes. We examine the effects of triggers targeting four ribosomal proteins (RP) transcripts. Although no significant mortality was observed after dsRNA microinjection and subsequent blood feeding, significant contrasts were observed on fecundity. Triggers targeting RPS6 and RPL26 effectively reduced gene expression but more importantly, reduced reproductive output by more than 96% and 91% at the first oviposition while triggers targeting RPL1 and RPS2 did not cause a reduction although gene expression was reduced. Significantly reduced fecundity continued through a second oviposition cycle in dsRPS6 and dsRPL26 cohorts, although the effect was not as strong. Relative gene expression levels confirmed specific transcript knockdown up to 20 days post-injection in mosquitoes that did not oviposit or produced reduced clutch sizes. Dissections at 36 h post-blood meal indicated defects in oocyte provisioning. The strong phenotype produced by dsRPS6 allowed us to examine the effects in various tissues as well as the dose response, trigger format, delivery method and trigger specificity in Aedes aegypti. Strong knockdown was observed in the abdomen and the ovaries. Greater than 50 ng of dsRPS6 significantly reduced fecundity but not when delivered in a sugar meal or as an siRNA. Similar bioassays with mutated dsRPS6 triggers indicates that up to three mismatches per possible siRNA are still effective in reducing fecundity. These studies indicate that while active and effective triggers can be developed for vector species, the lack of an efficient delivery method is the biggest barrier to use as a potential control method.

View all citing articles on Scopus

View full text

Article preview

Biochemical and Biophysical Research Communications

Abstract

Section snippets

Materials and methods

Results and discussion

Acknowledgments

References (24)

Tick-borne bacterial diseases emerging in Europe

Clin. Microbiol. Infect.

The ecological basis of parasite control: ticks and flies

Vet. Parasitol.

Identification of protective antigens for the control of Ixodes scapularis infestations using cDNA expression library immunization

Vaccine

Characterization of three Ixodes scapularis cDNAs protective against tick infestations

Vaccine

Vaccination with recombinant tick antigens for the control of Ixodes scapularis adult infestations

Vaccine

Systematics and evolution of ticks with a list of valid genus and species names

Parasitology

Nutrition and course of spermatogenesis in male ixodids

Dokl. Akad. Nauk. USSR

Ticks feeding on humans: a review of records on human-biting Ixodoidea with special reference to pathogen transmission

Exp. Appl. Acarol.

Anaplasma marginale (Rickettsiales: Anaplasmataceae): recent advances in defining host-pathogen adaptations of a tick-borne rickettsia

Parasitology

Advances in the identification and characterization of protective antigens for development of recombinant vaccines against tick infestations

Expert Rev. Vaccines

Anti-tick vaccines

Parasitology

Population genetics of autocidal control and strain replacement

Annu. Rev. Entomol.

Cited by (58)

Innovative approaches for the control of ticks and tick-borne diseases

Control of tick infestations in wild roe deer (Capreolus capreolus) vaccinated with the Q38 Subolesin/Akirin chimera

Controlling ticks and tick-borne diseases…looking forward

Liposome mediated double-stranded RNA delivery to silence ribosomal protein P0 in the tick Rhipicephalus haemaphysaloides

Targeting a global health problem: Vaccine design and challenges for the control of tick-borne diseases

DsRNA-mediated targeting of ribosomal transcripts RPS6 and RPL26 induces long-lasting and significant reductions in fecundity of the vector Aedes aegypti