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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...
Autores principales: | , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2003
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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. |
format | Text |
id | pubmed-149360 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2003 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
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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