Genetics and Biochemistry of RNAi in Drosophila

Kavi, Harsh H.; Fernandez, Harvey R.; Xie, Weiwu; Birchler, James A.

doi:10.1007/978-3-540-75157-1_3

Cited by 12 publications

(9 citation statements)

References 171 publications

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“…The former is based on the technique of RNA interference (RNAi) that was first discovered in Caenorhabditis elegans [ 6 ]. RNAi is a gene silencing mechanism in which a double-stranded RNA (dsRNA) molecule is generated that directs the specific degradation of the corresponding target mRNA [ 34 ]. The BDNF-antisense-GFP amplicon targets and degrades the cytoplasmic mRNA pool of BDNF through the mechanism of RNA interference: expression of the transgene leads to formation of sense RNA-RNA hybrids in the cytosol and thereby degrades and/or prevents translation of pro-BDNF mRNA.…”

Section: Discussionmentioning

confidence: 99%

Silencing Status Epilepticus-Induced BDNF Expression with Herpes Simplex Virus Type-1 Based Amplicon Vectors

et al. 2016

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Brain-derived neurotrophic factor (BDNF) has been found to produce pro- but also anti-epileptic effects. Thus, its validity as a therapeutic target must be verified using advanced tools designed to block or to enhance its signal. The aim of this study was to develop tools to silence the BDNF signal. We generated Herpes simplex virus type 1 (HSV-1) derived amplicon vectors, i.e. viral particles containing a genome of 152 kb constituted of concatameric repetitions of an expression cassette, enabling the expression of the gene of interest in multiple copies. HSV-1 based amplicon vectors are non-pathogenic and have been successfully employed in the past for gene delivery into the brain of living animals. Therefore, amplicon vectors should represent a logical choice for expressing a silencing cassette, which, in multiple copies, is expected to lead to an efficient knock-down of the target gene expression. Here, we employed two amplicon-based BDNF silencing strategies. The first, antisense, has been chosen to target and degrade the cytoplasmic mRNA pool of BDNF, whereas the second, based on the convergent transcription technology, has been chosen to repress transcription at the BDNF gene. Both these amplicon vectors proved to be effective in down-regulating BDNF expression in vitro, in BDNF-expressing mesoangioblast cells. However, only the antisense strategy was effective in vivo, after inoculation in the hippocampus in a model of status epilepticus in which BDNF mRNA levels are strongly increased. Interestingly, the knocking down of BDNF levels induced with BDNF-antisense was sufficient to produce significant behavioral effects, in spite of the fact that it was produced only in a part of a single hippocampus. In conclusion, this study demonstrates a reliable effect of amplicon vectors in knocking down gene expression in vitro and in vivo. Therefore, this approach may find broad applications in neurobiological studies.

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Section: Discussionmentioning

confidence: 99%

Silencing Status Epilepticus-Induced BDNF Expression with Herpes Simplex Virus Type-1 Based Amplicon Vectors

et al. 2016

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“…Work from many laboratories has shown that the RNAi system is present in Drosophila and plays several important roles via posttranscriptional gene silencing. For example, there is a Dicer-1 dependent miRNA system impacting developmental regulation by messenger RNA degradation or translational inhibition, and a Dicer-2 dependent small interfering RNA (siRNA) system active in viral defense (reviewed in Kavi et al 2008;Huisinga and Elgin 2009).…”

Section: How Is Heterochromatin Formation Targeted In Drosophila?mentioning

confidence: 99%

Position-Effect Variegation, Heterochromatin Formation, and Gene Silencing in Drosophila

Elgin

Reuter

2013

Cold Spring Harbor Perspectives in Biology

452

470

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“…Drosophila is an excellent model organism for this research because of the availability of its complete, assembled, well-annotated genome sequence [ 10 , 11 ] and genetics systems for facile induction and analysis of mutations. For comprehensive reviews of Drosophila RNAi see [ 12 , 13 , 14 ].…”

Section: The Exo-sirna Pathway Is An Antiviral Defense Mechanism Imentioning

confidence: 99%

The Role of RNA Interference (RNAi) in Arbovirus-Vector Interactions

Blair

Olson

2015

Viruses

140

132

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RNA interference (RNAi) was shown over 18 years ago to be a mechanism by which arbovirus replication and transmission could be controlled in arthropod vectors. During the intervening period, research on RNAi has defined many of the components and mechanisms of this antiviral pathway in arthropods, yet a number of unexplored questions remain. RNAi refers to RNA-mediated regulation of gene expression. Originally, the term described silencing of endogenous genes by introduction of exogenous double-stranded (ds)RNA with the same sequence as the gene to be silenced. Further research has shown that RNAi comprises three gene regulation pathways that are mediated by small RNAs: the small interfering (si)RNA, micro (mi)RNA, and Piwi-interacting (pi)RNA pathways. The exogenous (exo-)siRNA pathway is now recognized as a major antiviral innate immune response of arthropods. More recent studies suggest that the piRNA and miRNA pathways might also have important roles in arbovirus-vector interactions. This review will focus on current knowledge of the role of the exo-siRNA pathway as an arthropod vector antiviral response and on emerging research into vector piRNA and miRNA pathway modulation of arbovirus-vector interactions. Although it is assumed that arboviruses must evade the vector’s antiviral RNAi response in order to maintain their natural transmission cycles, the strategies by which this is accomplished are not well defined. RNAi is also an important tool for arthropod gene knock-down in functional genomics studies and in development of arbovirus-resistant mosquito populations. Possible arbovirus strategies for evasion of RNAi and applications of RNAi in functional genomics analysis and arbovirus transmission control will also be reviewed.

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Genetics and Biochemistry of RNAi in Drosophila

Cited by 12 publications

References 171 publications

Silencing Status Epilepticus-Induced BDNF Expression with Herpes Simplex Virus Type-1 Based Amplicon Vectors

Silencing Status Epilepticus-Induced BDNF Expression with Herpes Simplex Virus Type-1 Based Amplicon Vectors

Position-Effect Variegation, Heterochromatin Formation, and Gene Silencing in Drosophila

The Role of RNA Interference (RNAi) in Arbovirus-Vector Interactions

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