A Comparative Analysis of RNAi Trigger Uptake and Distribution in Mosquito Vectors of Disease
In mosquitoes, the utilization of RNAi for functional genetics is widespread, usually mediated through introduced double-stranded RNAs (dsRNAs) with sequence identity to a gene of interest. However, RNAi in mosquitoes is often hampered by inconsistencies in target gene knockdown between experimental...
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| Format: | Article |
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MDPI AG
2023-06-01
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| Series: | Insects |
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| Online Access: | https://www.mdpi.com/2075-4450/14/6/556 |
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| author | Paul M. Airs Katherine E. Kudrna Bailey Lubinski Yashdeep Phanse Lyric C. Bartholomay |
| author_facet | Paul M. Airs Katherine E. Kudrna Bailey Lubinski Yashdeep Phanse Lyric C. Bartholomay |
| author_sort | Paul M. Airs |
| collection | DOAJ |
| description | In mosquitoes, the utilization of RNAi for functional genetics is widespread, usually mediated through introduced double-stranded RNAs (dsRNAs) with sequence identity to a gene of interest. However, RNAi in mosquitoes is often hampered by inconsistencies in target gene knockdown between experimental setups. While the core RNAi pathway is known to function in most mosquito strains, the uptake and biodistribution of dsRNAs across different mosquito species and life stages have yet to be extensively explored as a source of variation in RNAi experiments. To better understand mosquito-RNAi dynamics, the biodistribution of a dsRNA to a heterologous gene, LacZ (iLacZ), was tracked following various routes of exposure in the larval and adult stages of <i>Aedes aegypti</i>, <i>Anopheles gambiae</i>, and <i>Culex pipiens</i>. iLacZ was largely limited to the gut lumen when exposed per os, or to the cuticle when topically applied, but spread through the hemocoel when injected. Uptake of dsRNA was noted in a subset of cells including: hemocytes, pericardial cells of the dorsal vessel, ovarian follicles, and ganglia of the ventral nerve cord. These cell types are all known to undergo phagocytosis, pinocytosis, or both, and as such may actively take up RNAi triggers. In <i>Ae. aegypti</i>, iLacZ was detected for up to one week post exposure by Northern blotting, but uptake and degradation drastically differed across tissues. The results presented here reveal that the uptake of RNAi triggers is distinct and specific to the cell type in vivo. |
| first_indexed | 2024-03-11T02:19:58Z |
| format | Article |
| id | doaj.art-ff624c96f6354afc9df67e511aefaeda |
| institution | Directory Open Access Journal |
| issn | 2075-4450 |
| language | English |
| last_indexed | 2024-03-11T02:19:58Z |
| publishDate | 2023-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Insects |
| spelling | doaj.art-ff624c96f6354afc9df67e511aefaeda2023-11-18T10:56:26ZengMDPI AGInsects2075-44502023-06-0114655610.3390/insects14060556A Comparative Analysis of RNAi Trigger Uptake and Distribution in Mosquito Vectors of DiseasePaul M. Airs0Katherine E. Kudrna1Bailey Lubinski2Yashdeep Phanse3Lyric C. Bartholomay4Department of Pathobiological Sciences, University of Wisconsin—Madison, Madison, WI 53706, USADepartment of Pathobiological Sciences, University of Wisconsin—Madison, Madison, WI 53706, USADepartment of Pathobiological Sciences, University of Wisconsin—Madison, Madison, WI 53706, USADepartment of Pathobiological Sciences, University of Wisconsin—Madison, Madison, WI 53706, USADepartment of Pathobiological Sciences, University of Wisconsin—Madison, Madison, WI 53706, USAIn mosquitoes, the utilization of RNAi for functional genetics is widespread, usually mediated through introduced double-stranded RNAs (dsRNAs) with sequence identity to a gene of interest. However, RNAi in mosquitoes is often hampered by inconsistencies in target gene knockdown between experimental setups. While the core RNAi pathway is known to function in most mosquito strains, the uptake and biodistribution of dsRNAs across different mosquito species and life stages have yet to be extensively explored as a source of variation in RNAi experiments. To better understand mosquito-RNAi dynamics, the biodistribution of a dsRNA to a heterologous gene, LacZ (iLacZ), was tracked following various routes of exposure in the larval and adult stages of <i>Aedes aegypti</i>, <i>Anopheles gambiae</i>, and <i>Culex pipiens</i>. iLacZ was largely limited to the gut lumen when exposed per os, or to the cuticle when topically applied, but spread through the hemocoel when injected. Uptake of dsRNA was noted in a subset of cells including: hemocytes, pericardial cells of the dorsal vessel, ovarian follicles, and ganglia of the ventral nerve cord. These cell types are all known to undergo phagocytosis, pinocytosis, or both, and as such may actively take up RNAi triggers. In <i>Ae. aegypti</i>, iLacZ was detected for up to one week post exposure by Northern blotting, but uptake and degradation drastically differed across tissues. The results presented here reveal that the uptake of RNAi triggers is distinct and specific to the cell type in vivo.https://www.mdpi.com/2075-4450/14/6/556RNA interferencedsRNAbiodistributionper os<i>Aedes</i><i>Anopheles</i> |
| spellingShingle | Paul M. Airs Katherine E. Kudrna Bailey Lubinski Yashdeep Phanse Lyric C. Bartholomay A Comparative Analysis of RNAi Trigger Uptake and Distribution in Mosquito Vectors of Disease Insects RNA interference dsRNA biodistribution per os <i>Aedes</i> <i>Anopheles</i> |
| title | A Comparative Analysis of RNAi Trigger Uptake and Distribution in Mosquito Vectors of Disease |
| title_full | A Comparative Analysis of RNAi Trigger Uptake and Distribution in Mosquito Vectors of Disease |
| title_fullStr | A Comparative Analysis of RNAi Trigger Uptake and Distribution in Mosquito Vectors of Disease |
| title_full_unstemmed | A Comparative Analysis of RNAi Trigger Uptake and Distribution in Mosquito Vectors of Disease |
| title_short | A Comparative Analysis of RNAi Trigger Uptake and Distribution in Mosquito Vectors of Disease |
| title_sort | comparative analysis of rnai trigger uptake and distribution in mosquito vectors of disease |
| topic | RNA interference dsRNA biodistribution per os <i>Aedes</i> <i>Anopheles</i> |
| url | https://www.mdpi.com/2075-4450/14/6/556 |
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