Mosquito tagging using DNA-barcoded nanoporous protein microcrystals

Julius D Stuart; Daniel A Hartman; Lyndsey I Gray; Alec A Jones; Natalie R Wickenkamp; Christine Hirt; Aya Safira; April R Regas; Therese M Kondash; Margaret L Yates; Sergei Driga; Christopher D Snow; Rebekah C Kading

doi:10.1093/pnasnexus/pgac190

Mosquito tagging using DNA-barcoded nanoporous protein microcrystals

PNAS Nexus. 2022 Sep 12;1(4):pgac190. doi: 10.1093/pnasnexus/pgac190. eCollection 2022 Sep.

Authors

Julius D Stuart¹, Daniel A Hartman^{2

3}, Lyndsey I Gray², Alec A Jones⁴, Natalie R Wickenkamp², Christine Hirt², Aya Safira⁴, April R Regas⁵, Therese M Kondash^{6

7}, Margaret L Yates⁸, Sergei Driga⁹, Christopher D Snow^{1

4

8

9}, Rebekah C Kading

Affiliations

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 Department of Entomology, Cornell University, Ithaca, NY 14853, USA.
⁴ School of Biomedical Engineering, Colorado State University, Fort Collins, CO 80523, USA.
⁵ College of Veterinary Medicine and Biological Sciences, Colorado State University, Fort Collins, CO 80523, USA.
⁶ Department of Environmental Health and Radiological Sciences, Colorado State University, Fort Collins, CO 80523, USA.
⁷ H3 Environmental, Albuquerque, NM 87109 (current).
⁸ Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, CO 80523, USA.
⁹ Department of Chemical and Biological Engineering, Colorado State University, Fort Collins, Colorado 80523, USA.

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.

Keywords: host-guest crystals; mark–release–recapture; mosquito surveillance; porous protein crystals; vector biology.