Abstract

Conventional mosquito marking technology for mark–release–recapture (MRR) is quite limited in terms of information capacity and efficacy. To overcome both challenges, we have engineered, lab-tested, and field-evaluated a new class of marker particles, in which synthetic, short DNA oligonucleotides (DNA barcodes) are adsorbed and protected within tough, crosslinked porous protein microcrystals. Mosquitoes self-mark through ingestion of microcrystals in their larval habitat. Barcoded microcrystals persist trans-stadially through mosquito development if ingested by larvae, do not significantly affect adult mosquito survivorship, and individual barcoded mosquitoes are detectable in pools of up to at least 20 mosquitoes. We have also demonstrated crystal persistence following adult mosquito ingestion. Barcode sequences can be recovered by qPCR and next-generation sequencing (NGS) without detectable amplification of native mosquito DNA. These DNA-laden protein microcrystals have the potential to radically increase the amount of information obtained from future MRR studies compared to previous studies employing conventional mosquito marking materials.

Details

Title
Mosquito tagging using DNA-barcoded nanoporous protein microcrystals
Author
Stuart, Julius D 1   VIAFID ORCID Logo  ; Hartman, Daniel A 2   VIAFID ORCID Logo  ; Gray, Lyndsey I 2   VIAFID ORCID Logo  ; Jones, Alec A 3   VIAFID ORCID Logo  ; Wickenkamp, Natalie R 2   VIAFID ORCID Logo  ; Hirt, Christine 2 ; Safira, Aya 3 ; Regas, April R 4   VIAFID ORCID Logo  ; Kondash, Therese M 5   VIAFID ORCID Logo  ; Yates, Margaret L 6   VIAFID ORCID Logo  ; Driga, Sergei 7 ; Snow, Christopher D 1   VIAFID ORCID Logo  ; Kading, Rebekah C  VIAFID ORCID Logo 

1  Department of Chemistry, Colorado State University , Fort Collins, CO 80523, USA 
2  Department of Microbiology, Immunology, and Pathology, Colorado State University , Fort Collins, CO 80523, USA 
3  School of Biomedical Engineering, Colorado State University , Fort Collins, CO 80523, USA 
4  College of Veterinary Medicine and Biological Sciences, Colorado State University , Fort Collins, CO 80523, USA 
5  Department of Environmental Health and Radiological Sciences, Colorado State University , Fort Collins, CO 80523, USA 
6  Department of Biochemistry and Molecular Biology, Colorado State University , Fort Collins, CO 80523, USA 
7  Department of Chemical and Biological Engineering, Colorado State University , Fort Collins, Colorado 80523, USA 
Publication year
2022
Publication date
Sep 2022
Publisher
Oxford University Press
e-ISSN
27526542
Source type
Scholarly Journal
Language of publication
English
DOI
https://doi.org/10.1093/pnasnexus/pgac190
ProQuest document ID
3191367381
Copyright
© The Author(s) 2022. Published by Oxford University Press on behalf of National Academy of Sciences. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.