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Comprehensive characterization of a transgene insertion in a highly repetitive, centromeric region of Anopheles mosquitoes
The availability of the genomic sequence of the malaria mosquito Anopheles gambiae has in recent years sparked the development of transgenic technologies with the potential to be used as novel vector control tools. These technologies rely on genome editing that confer traits able to affect vectorial...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Taylor & Francis
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10081084/ https://www.ncbi.nlm.nih.gov/pubmed/35861105 http://dx.doi.org/10.1080/20477724.2022.2100192 |
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author | Vitale, Matteo Leo, Chiara Courty, Thomas Kranjc, Nace Connolly, John B. Morselli, Giulia Bamikole, Christopher Haghighat-Khah, Roya Elaine Bernardini, Federica Fuchs, Silke |
author_facet | Vitale, Matteo Leo, Chiara Courty, Thomas Kranjc, Nace Connolly, John B. Morselli, Giulia Bamikole, Christopher Haghighat-Khah, Roya Elaine Bernardini, Federica Fuchs, Silke |
author_sort | Vitale, Matteo |
collection | PubMed |
description | The availability of the genomic sequence of the malaria mosquito Anopheles gambiae has in recent years sparked the development of transgenic technologies with the potential to be used as novel vector control tools. These technologies rely on genome editing that confer traits able to affect vectorial capacity. This can be achieved by either reducing the mosquito population or by making mosquitoes refractory to the parasite infection. For any genetically modified organism that is regarded for release, molecular characterization of the transgene and flanking sites are essential for their safety assessment and post-release monitoring. Despite great advancements, Whole-Genome Sequencing data are still subject to limitations due to the presence of repetitive and unannotated DNA sequences. Faced with this challenge, we describe a number of techniques that were used to identify the genomic location of a transgene in the male bias mosquito strain Ag(PMB)1 considered for potential field application. While the initial inverse PCR identified the most likely insertion site on Chromosome 3 R 36D, reassessment of the data showed a high repetitiveness in those sequences and multiple genomic locations as potential insertion sites of the transgene. Here we used a combination of DNA sequencing analysis and in-situ hybridization to clearly identify the integration of the transgene in a poorly annotated centromeric region of Chromosome 2 R 19D. This study emphasizes the need for accuracy in sequencing data for the genome of organisms of medical importance such as Anopheles mosquitoes and other tools available that can support genomic locations of transgenes. |
format | Online Article Text |
id | pubmed-10081084 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Taylor & Francis |
record_format | MEDLINE/PubMed |
spelling | pubmed-100810842023-04-08 Comprehensive characterization of a transgene insertion in a highly repetitive, centromeric region of Anopheles mosquitoes Vitale, Matteo Leo, Chiara Courty, Thomas Kranjc, Nace Connolly, John B. Morselli, Giulia Bamikole, Christopher Haghighat-Khah, Roya Elaine Bernardini, Federica Fuchs, Silke Pathog Glob Health Articles The availability of the genomic sequence of the malaria mosquito Anopheles gambiae has in recent years sparked the development of transgenic technologies with the potential to be used as novel vector control tools. These technologies rely on genome editing that confer traits able to affect vectorial capacity. This can be achieved by either reducing the mosquito population or by making mosquitoes refractory to the parasite infection. For any genetically modified organism that is regarded for release, molecular characterization of the transgene and flanking sites are essential for their safety assessment and post-release monitoring. Despite great advancements, Whole-Genome Sequencing data are still subject to limitations due to the presence of repetitive and unannotated DNA sequences. Faced with this challenge, we describe a number of techniques that were used to identify the genomic location of a transgene in the male bias mosquito strain Ag(PMB)1 considered for potential field application. While the initial inverse PCR identified the most likely insertion site on Chromosome 3 R 36D, reassessment of the data showed a high repetitiveness in those sequences and multiple genomic locations as potential insertion sites of the transgene. Here we used a combination of DNA sequencing analysis and in-situ hybridization to clearly identify the integration of the transgene in a poorly annotated centromeric region of Chromosome 2 R 19D. This study emphasizes the need for accuracy in sequencing data for the genome of organisms of medical importance such as Anopheles mosquitoes and other tools available that can support genomic locations of transgenes. Taylor & Francis 2022-07-21 /pmc/articles/PMC10081084/ /pubmed/35861105 http://dx.doi.org/10.1080/20477724.2022.2100192 Text en © 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Articles Vitale, Matteo Leo, Chiara Courty, Thomas Kranjc, Nace Connolly, John B. Morselli, Giulia Bamikole, Christopher Haghighat-Khah, Roya Elaine Bernardini, Federica Fuchs, Silke Comprehensive characterization of a transgene insertion in a highly repetitive, centromeric region of Anopheles mosquitoes |
title | Comprehensive characterization of a transgene insertion in a highly repetitive, centromeric region of Anopheles mosquitoes |
title_full | Comprehensive characterization of a transgene insertion in a highly repetitive, centromeric region of Anopheles mosquitoes |
title_fullStr | Comprehensive characterization of a transgene insertion in a highly repetitive, centromeric region of Anopheles mosquitoes |
title_full_unstemmed | Comprehensive characterization of a transgene insertion in a highly repetitive, centromeric region of Anopheles mosquitoes |
title_short | Comprehensive characterization of a transgene insertion in a highly repetitive, centromeric region of Anopheles mosquitoes |
title_sort | comprehensive characterization of a transgene insertion in a highly repetitive, centromeric region of anopheles mosquitoes |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10081084/ https://www.ncbi.nlm.nih.gov/pubmed/35861105 http://dx.doi.org/10.1080/20477724.2022.2100192 |
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