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Disruption of vitellogenin gene function in adult honeybees by intra-abdominal injection of double-stranded RNA

BACKGROUND: The ability to manipulate the genetic networks underlying the physiological and behavioural repertoires of the adult honeybee worker (Apis mellifera) is likely to deepen our understanding of issues such as learning and memory generation, ageing, and the regulatory anatomy of social syste...

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Autores principales: Amdam, Gro V, Simões, Zilá LP, Guidugli, Karina R, Norberg, Kari, Omholt, Stig W
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2003
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC149360/
https://www.ncbi.nlm.nih.gov/pubmed/12546706
http://dx.doi.org/10.1186/1472-6750-3-1
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author Amdam, Gro V
Simões, Zilá LP
Guidugli, Karina R
Norberg, Kari
Omholt, Stig W
author_facet Amdam, Gro V
Simões, Zilá LP
Guidugli, Karina R
Norberg, Kari
Omholt, Stig W
author_sort Amdam, Gro V
collection PubMed
description BACKGROUND: The ability to manipulate the genetic networks underlying the physiological and behavioural repertoires of the adult honeybee worker (Apis mellifera) is likely to deepen our understanding of issues such as learning and memory generation, ageing, and the regulatory anatomy of social systems in proximate as well as evolutionary terms. Here we assess two methods for probing gene function by RNA interference (RNAi) in adult honeybees. RESULTS: The vitellogenin gene was chosen as target because its expression is unlikely to have a phenotypic effect until the adult stage in bees. This allowed us to introduce dsRNA in preblastoderm eggs without affecting gene function during development. Of workers reared from eggs injected with dsRNA derived from a 504 bp stretch of the vitellogenin coding sequence, 15% had strongly reduced levels of vitellogenin mRNA. When dsRNA was introduced by intra-abdominal injection in newly emerged bees, almost all individuals (96 %) showed the mutant phenotype. An RNA-fragment with an apparent size similar to the template dsRNA was still present in this group after 15 days. CONCLUSION: Injection of dsRNA in eggs at the preblastoderm stage seems to allow disruption of gene function in all developmental stages. To dissect gene function in the adult stage, the intra-abdominal injection technique seems superior to egg injection as it gives a much higher penetrance, it is much simpler, and it makes it possible to address genes that are also expressed in the embryonic, larval or pupal stages.
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spelling pubmed-1493602003-02-25 Disruption of vitellogenin gene function in adult honeybees by intra-abdominal injection of double-stranded RNA Amdam, Gro V Simões, Zilá LP Guidugli, Karina R Norberg, Kari Omholt, Stig W BMC Biotechnol Methodology Article BACKGROUND: The ability to manipulate the genetic networks underlying the physiological and behavioural repertoires of the adult honeybee worker (Apis mellifera) is likely to deepen our understanding of issues such as learning and memory generation, ageing, and the regulatory anatomy of social systems in proximate as well as evolutionary terms. Here we assess two methods for probing gene function by RNA interference (RNAi) in adult honeybees. RESULTS: The vitellogenin gene was chosen as target because its expression is unlikely to have a phenotypic effect until the adult stage in bees. This allowed us to introduce dsRNA in preblastoderm eggs without affecting gene function during development. Of workers reared from eggs injected with dsRNA derived from a 504 bp stretch of the vitellogenin coding sequence, 15% had strongly reduced levels of vitellogenin mRNA. When dsRNA was introduced by intra-abdominal injection in newly emerged bees, almost all individuals (96 %) showed the mutant phenotype. An RNA-fragment with an apparent size similar to the template dsRNA was still present in this group after 15 days. CONCLUSION: Injection of dsRNA in eggs at the preblastoderm stage seems to allow disruption of gene function in all developmental stages. To dissect gene function in the adult stage, the intra-abdominal injection technique seems superior to egg injection as it gives a much higher penetrance, it is much simpler, and it makes it possible to address genes that are also expressed in the embryonic, larval or pupal stages. BioMed Central 2003-01-20 /pmc/articles/PMC149360/ /pubmed/12546706 http://dx.doi.org/10.1186/1472-6750-3-1 Text en Copyright © 2003 Amdam et al; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.
spellingShingle Methodology Article
Amdam, Gro V
Simões, Zilá LP
Guidugli, Karina R
Norberg, Kari
Omholt, Stig W
Disruption of vitellogenin gene function in adult honeybees by intra-abdominal injection of double-stranded RNA
title Disruption of vitellogenin gene function in adult honeybees by intra-abdominal injection of double-stranded RNA
title_full Disruption of vitellogenin gene function in adult honeybees by intra-abdominal injection of double-stranded RNA
title_fullStr Disruption of vitellogenin gene function in adult honeybees by intra-abdominal injection of double-stranded RNA
title_full_unstemmed Disruption of vitellogenin gene function in adult honeybees by intra-abdominal injection of double-stranded RNA
title_short Disruption of vitellogenin gene function in adult honeybees by intra-abdominal injection of double-stranded RNA
title_sort disruption of vitellogenin gene function in adult honeybees by intra-abdominal injection of double-stranded rna
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC149360/
https://www.ncbi.nlm.nih.gov/pubmed/12546706
http://dx.doi.org/10.1186/1472-6750-3-1
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