RNA interference to reveal roles ofβ-N-acetylglucosaminidase gene during molting process inLocusta migratoria

2012 ◽  
Vol 20 (1) ◽  
pp. 109-119 ◽  
Author(s):  
Shuo Rong ◽  
Da-Qi Li ◽  
Xue-Yao Zhang ◽  
Sheng Li ◽  
Kun Yan Zhu ◽  
...  
Keyword(s):  
Rna Interference ◽  
Molting Process

Regulatory functions of trehalose-6-phosphate synthase in the chitin biosynthesis pathway in Tribolium castaneum (Coleoptera: Tenebrionidae) revealed by RNA interference

2017 ◽  
Vol 108 (3) ◽  
pp. 388-399 ◽  
Author(s):  
Q.W. Chen ◽  
S. Jin ◽  
L. Zhang ◽  
Q.D. Shen ◽  
P. Wei ◽  
...  
Keyword(s):  
Rna Interference ◽  
Tribolium Castaneum ◽  
Fat Body ◽  
Regulation Of Gene Expression ◽  
Chitin Synthesis ◽  
Reading Frame ◽  
Molting Process ◽  
Polymerase Chain ◽  
Regulatory Functions ◽  
Phosphate Synthase

AbstractRNA interference (RNAi) is a very effective technique for studying gene function and may be an efficient method for controlling pests. Trehalose-6-phosphate synthase (TPS), which plays a key role in the synthesis of trehalose and insect development, was cloned in Tribolium castaneum (Herbst) (TcTPS) and the putative functions were studied using RNAi via the injection of double-stranded RNA (dsRNA) corresponding to conserved TPS and trehalose-6-phosphate phosphatase domains. Expression analyses show that TcTPS is expressed higher in the fat body, while quantitative real-time polymerase chain reaction results show that the expression of four trehalase isoforms was significantly suppressed by dsTPS injection. Additionally, the expression of six chitin synthesis-related genes, such as hexokinase 2 and glutamine-fructose-6-phosphate aminotransferase, was suppressed at 48 and 72 h post-dsTPS-1 and dsTPS-2 RNA injection, which were two dsTPS fragments that had been designed for two different locations in TcTPS open reading frame, and that trehalose content and trehalase 1 activity decreased significantly at 72 h post-dsRNA injection. Furthermore, T. castaneum injected with dsTPS-1 and dsTPS-2 RNA displayed significantly lower levels of chitin and could not complete the molting process from larvae to pupae, revealing abnormal molting phenotypes. These results demonstrate that silencing TPS gene leads to molting deformities and high mortality rates via regulation of gene expression in the chitin biosynthetic pathway, and may be a promising approach for pest control in the future.

scholarly journals Pyrophosphatase of the Roundworm Ascaris suum Plays an Essential Role in the Worm's Molting and Development

2005 ◽  
Vol 73 (4) ◽  
pp. 1995-2004 ◽  
Author(s):  
M. Khyrul Islam ◽  
Takeharu Miyoshi ◽  
Manabu Yamada ◽  
Naotoshi Tsuji
Keyword(s):  
Rna Interference ◽  
Active Sites ◽  
Ascaris Suum ◽  
Critical Role ◽  
Interleukin 2 ◽  
Pcr Analysis ◽  
Mrna Levels ◽  
Molting Process ◽  
Third Stage

ABSTRACT Previous studies indicated that inorganic pyrophosphatase of Ascaris suum (AsPPase) plays an important role in larval survival in the host. Here we describe a precise role for AsPPase in larval molting and development and also describe the potential role of recombinant AsPPase (rAsPPase) in protective immunity to A. suum infection. Using reverse transcriptase PCR analysis, we found that disruption of AsPPase gene function by RNA interference resulted in suppression of AsPPase mRNA levels. RNA interference also caused inhibition of molting of third-stage larvae (31%) and suppression of native protein expression, as demonstrated by a 56% reduction in enzyme activity and quantified by immunoblot and immunofluorescence analyses, suggesting that AsPPase has a role in the molting process. The anatomic location of the AsPPase native enzyme in the hypodermis of larvae along with its elevated expression prior to and during the molting process supports such a role. Anti-rAsPPase immunoglobulin G (IgG) also resulted in 57% inhibition of molting of A. suum lung-stage third-stage larvae to fourth-stage larvae in vitro with developmental arrest. Antigenic epitopes of AsPPase overlapped the enzyme active sites. Mice immunized with rAsPPase exhibited high antigen-specific IgG antibody responses and were protected (>70%) against a challenge A. suum migratory-phase infection. Splenic T cells from rAsPPase-immunized mice produced low levels of T helper 1-type cytokines (gamma interferon and interleukin-2) in vitro but exhibited an elevated interleukin-10 response. A significantly high level of IgG1 subclass antibodies was found in immunized mice. Our results establish that AsPPase has a critical role in the molting and development of Ascaris roundworms and suggest the potential of AsPPase for use as a candidate vaccine against ascariasis.

RNA Interference Technology

2005 ◽  
Author(s):  
Andrew Fire ◽  
Marshall Nirenberg
Keyword(s):  
Rna Interference

The oncolytic adenovirus Ad-iREP-2 utilizes p53-dependent rna interference for tumor-selective replication

2008 ◽  
Vol 46 (09) ◽  
Author(s):  
E Gürlevik ◽  
P Schache ◽  
L Zender ◽  
MP Manns ◽  
S Kubicka ◽  
...  
Keyword(s):  
Rna Interference ◽  
Oncolytic Adenovirus ◽  
Tumor Selective

RNA interference and its clinical applications

2007 ◽  
Vol 148 (47) ◽  
pp. 2235-2240 ◽  
Author(s):  
Gyöngyi Munkácsy ◽  
Zsolt Tulassay ◽  
Balázs Győrffy
Keyword(s):  
Rna Interference ◽  
Clinical Applications

Az RNS-interferencia a poszttranszkripciós génelcsendesítés olyan formája, amelynek során rövid, specifikusan RNS-molekulák elnyomják a gének kifejeződésében kulcsszerepet játszó hírvivő RNS-ek működését. A sejtbe juttatott dupla szálú vagy rövid interferáló RNS-molekulák aktiválják az RNS-indukált elcsendesítő komplexet, amely a célgén hírvivő RNS-ét lebontja. A sejtek saját szabályozó mikro-RNS-molekulákkal is rendelkeznek, amelyeknek hírvivő RNS-e képes önmagával hajtűt képezni, amit a sejt dupla szálú RNS-ként értelmez. Az RNS-interferencia élettani működései közé tartozik a vírusok és a transzpozonok elleni védekezés, valamint a génkifejeződés szabályozása. Az RNS-interferencia nemcsak in vitro alkalmazható az egyes gének működésének vizsgálatára, hanem klinikai alkalmazásainak lehetőségei is megjelentek. Eddig vírusfertőzésekben, az időskori makuladegeneráció gátlására, a vér koleszterinszint-csökkentésére, daganatellenes és neurodegeneratív betegségek kezelésében alkalmazták. Az RNS-interferencia alkalmazását azonban nehezíti, hogy a megfelelő rövid interferáló RNS-molekulák tervezéséhez szükséges bioinformatikai algoritmusok nem tökéletesek; a szervezet szöveteibe való bejuttatásuk nehéz; illetve csak olyan esetekben alkalmazható, amelyekben átmeneti antagonista génelcsendesítő hatás és nem hosszú távú kezelés szükséges. Az alkalmazás legnagyobb előnye a jelentős specificitás, ami miatt mellékhatása is kevés. Az RNS-interferencia alapú kezelések megjelenése már a közeli jövőben várható.

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