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Methods for the generation of heritable germline mutations in the disease vector Culex quinquefasciatus using clustered regularly interspaced short palindrome repeats‐associated pr...
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| Zeitschriftentitel: | Insect Molecular Biology |
|---|---|
| Personen und Körperschaften: | , , , , , |
| In: | Insect Molecular Biology, 29, 2020, 2, S. 214-220 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
Wiley
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| Schlagwörter: |
| author_facet |
Li, M. Li, T. Liu, N. Raban, R. R. Wang, X. Akbari, O. S. Li, M. Li, T. Liu, N. Raban, R. R. Wang, X. Akbari, O. S. |
|---|---|
| author |
Li, M. Li, T. Liu, N. Raban, R. R. Wang, X. Akbari, O. S. |
| spellingShingle |
Li, M. Li, T. Liu, N. Raban, R. R. Wang, X. Akbari, O. S. Insect Molecular Biology Methods for the generation of heritable germline mutations in the disease vector Culex quinquefasciatus using clustered regularly interspaced short palindrome repeats‐associated protein 9 Insect Science Genetics Molecular Biology |
| author_sort |
li, m. |
| spelling |
Li, M. Li, T. Liu, N. Raban, R. R. Wang, X. Akbari, O. S. 0962-1075 1365-2583 Wiley Insect Science Genetics Molecular Biology http://dx.doi.org/10.1111/imb.12626 <jats:title>Abstract</jats:title><jats:p><jats:italic>Culex quinquefasciatus</jats:italic> is a vector of many diseases that adversely impact human and animal health; however, compared to other mosquito vectors limited genome engineering technologies have been characterized for this vector. Clustered regularly interspaced short palindrome repeats‐associated protein 9 (CRISPR‐Cas9) based technologies are a powerful tool for genome engineering and functional genetics and consequently have transformed genetic studies in many organisms. Our objective was to improve upon the limited technologies available for genome editing in <jats:italic>C. quinquefasciatus</jats:italic> to create a reproducible and straightforward method for CRISPR‐Cas9‐targeted mutagenesis in this vector. Here we describe methods to achieve high embryo survival and mutagenesis rates and we provide details on the injection supplies and procedures, embryo handling and guide RNA (gRNA) target designs. Through these efforts, we achieved embryo survival rates and germline mutagenesis rates that greatly exceed previously reported rates in this vector. This work is also the first to characterize the <jats:italic>white</jats:italic> gene marker in this species, which is a valuable phenotypic marker for future transgenesis or mutagenesis of this vector. Overall, these tools provide the framework for future functional genetic studies in this important disease vector and may support the development of future gene drive and genetic technologies that can be used to control this vector.</jats:p> Methods for the generation of heritable germline mutations in the disease vector <i>Culex quinquefasciatus</i> using clustered regularly interspaced short palindrome repeats‐associated protein 9 Insect Molecular Biology |
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| title |
Methods for the generation of heritable germline mutations in the disease vector Culex quinquefasciatus using clustered regularly interspaced short palindrome repeats‐associated protein 9 |
| title_unstemmed |
Methods for the generation of heritable germline mutations in the disease vector Culex quinquefasciatus using clustered regularly interspaced short palindrome repeats‐associated protein 9 |
| title_full |
Methods for the generation of heritable germline mutations in the disease vector Culex quinquefasciatus using clustered regularly interspaced short palindrome repeats‐associated protein 9 |
| title_fullStr |
Methods for the generation of heritable germline mutations in the disease vector Culex quinquefasciatus using clustered regularly interspaced short palindrome repeats‐associated protein 9 |
| title_full_unstemmed |
Methods for the generation of heritable germline mutations in the disease vector Culex quinquefasciatus using clustered regularly interspaced short palindrome repeats‐associated protein 9 |
| title_short |
Methods for the generation of heritable germline mutations in the disease vector Culex quinquefasciatus using clustered regularly interspaced short palindrome repeats‐associated protein 9 |
| title_sort |
methods for the generation of heritable germline mutations in the disease vector <i>culex quinquefasciatus</i> using clustered regularly interspaced short palindrome repeats‐associated protein 9 |
| topic |
Insect Science Genetics Molecular Biology |
| url |
http://dx.doi.org/10.1111/imb.12626 |
| publishDate |
2020 |
| physical |
214-220 |
| description |
<jats:title>Abstract</jats:title><jats:p><jats:italic>Culex quinquefasciatus</jats:italic> is a vector of many diseases that adversely impact human and animal health; however, compared to other mosquito vectors limited genome engineering technologies have been characterized for this vector. Clustered regularly interspaced short palindrome repeats‐associated protein 9 (CRISPR‐Cas9) based technologies are a powerful tool for genome engineering and functional genetics and consequently have transformed genetic studies in many organisms. Our objective was to improve upon the limited technologies available for genome editing in <jats:italic>C. quinquefasciatus</jats:italic> to create a reproducible and straightforward method for CRISPR‐Cas9‐targeted mutagenesis in this vector. Here we describe methods to achieve high embryo survival and mutagenesis rates and we provide details on the injection supplies and procedures, embryo handling and guide RNA (gRNA) target designs. Through these efforts, we achieved embryo survival rates and germline mutagenesis rates that greatly exceed previously reported rates in this vector. This work is also the first to characterize the <jats:italic>white</jats:italic> gene marker in this species, which is a valuable phenotypic marker for future transgenesis or mutagenesis of this vector. Overall, these tools provide the framework for future functional genetic studies in this important disease vector and may support the development of future gene drive and genetic technologies that can be used to control this vector.</jats:p> |
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| author | Li, M., Li, T., Liu, N., Raban, R. R., Wang, X., Akbari, O. S. |
| author_facet | Li, M., Li, T., Liu, N., Raban, R. R., Wang, X., Akbari, O. S., Li, M., Li, T., Liu, N., Raban, R. R., Wang, X., Akbari, O. S. |
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| description | <jats:title>Abstract</jats:title><jats:p><jats:italic>Culex quinquefasciatus</jats:italic> is a vector of many diseases that adversely impact human and animal health; however, compared to other mosquito vectors limited genome engineering technologies have been characterized for this vector. Clustered regularly interspaced short palindrome repeats‐associated protein 9 (CRISPR‐Cas9) based technologies are a powerful tool for genome engineering and functional genetics and consequently have transformed genetic studies in many organisms. Our objective was to improve upon the limited technologies available for genome editing in <jats:italic>C. quinquefasciatus</jats:italic> to create a reproducible and straightforward method for CRISPR‐Cas9‐targeted mutagenesis in this vector. Here we describe methods to achieve high embryo survival and mutagenesis rates and we provide details on the injection supplies and procedures, embryo handling and guide RNA (gRNA) target designs. Through these efforts, we achieved embryo survival rates and germline mutagenesis rates that greatly exceed previously reported rates in this vector. This work is also the first to characterize the <jats:italic>white</jats:italic> gene marker in this species, which is a valuable phenotypic marker for future transgenesis or mutagenesis of this vector. Overall, these tools provide the framework for future functional genetic studies in this important disease vector and may support the development of future gene drive and genetic technologies that can be used to control this vector.</jats:p> |
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| spelling | Li, M. Li, T. Liu, N. Raban, R. R. Wang, X. Akbari, O. S. 0962-1075 1365-2583 Wiley Insect Science Genetics Molecular Biology http://dx.doi.org/10.1111/imb.12626 <jats:title>Abstract</jats:title><jats:p><jats:italic>Culex quinquefasciatus</jats:italic> is a vector of many diseases that adversely impact human and animal health; however, compared to other mosquito vectors limited genome engineering technologies have been characterized for this vector. Clustered regularly interspaced short palindrome repeats‐associated protein 9 (CRISPR‐Cas9) based technologies are a powerful tool for genome engineering and functional genetics and consequently have transformed genetic studies in many organisms. Our objective was to improve upon the limited technologies available for genome editing in <jats:italic>C. quinquefasciatus</jats:italic> to create a reproducible and straightforward method for CRISPR‐Cas9‐targeted mutagenesis in this vector. Here we describe methods to achieve high embryo survival and mutagenesis rates and we provide details on the injection supplies and procedures, embryo handling and guide RNA (gRNA) target designs. Through these efforts, we achieved embryo survival rates and germline mutagenesis rates that greatly exceed previously reported rates in this vector. This work is also the first to characterize the <jats:italic>white</jats:italic> gene marker in this species, which is a valuable phenotypic marker for future transgenesis or mutagenesis of this vector. Overall, these tools provide the framework for future functional genetic studies in this important disease vector and may support the development of future gene drive and genetic technologies that can be used to control this vector.</jats:p> Methods for the generation of heritable germline mutations in the disease vector <i>Culex quinquefasciatus</i> using clustered regularly interspaced short palindrome repeats‐associated protein 9 Insect Molecular Biology |
| spellingShingle | Li, M., Li, T., Liu, N., Raban, R. R., Wang, X., Akbari, O. S., Insect Molecular Biology, Methods for the generation of heritable germline mutations in the disease vector Culex quinquefasciatus using clustered regularly interspaced short palindrome repeats‐associated protein 9, Insect Science, Genetics, Molecular Biology |
| title | Methods for the generation of heritable germline mutations in the disease vector Culex quinquefasciatus using clustered regularly interspaced short palindrome repeats‐associated protein 9 |
| title_full | Methods for the generation of heritable germline mutations in the disease vector Culex quinquefasciatus using clustered regularly interspaced short palindrome repeats‐associated protein 9 |
| title_fullStr | Methods for the generation of heritable germline mutations in the disease vector Culex quinquefasciatus using clustered regularly interspaced short palindrome repeats‐associated protein 9 |
| title_full_unstemmed | Methods for the generation of heritable germline mutations in the disease vector Culex quinquefasciatus using clustered regularly interspaced short palindrome repeats‐associated protein 9 |
| title_short | Methods for the generation of heritable germline mutations in the disease vector Culex quinquefasciatus using clustered regularly interspaced short palindrome repeats‐associated protein 9 |
| title_sort | methods for the generation of heritable germline mutations in the disease vector <i>culex quinquefasciatus</i> using clustered regularly interspaced short palindrome repeats‐associated protein 9 |
| title_unstemmed | Methods for the generation of heritable germline mutations in the disease vector Culex quinquefasciatus using clustered regularly interspaced short palindrome repeats‐associated protein 9 |
| topic | Insect Science, Genetics, Molecular Biology |
| url | http://dx.doi.org/10.1111/imb.12626 |