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PreprintCC BY-NC-NDmedRxiv

Identification of genetic variants in Pfs25 and functional evaluation in mosquito infection

  • A. Orfano
  • A. Cisse
  • Y. Guo
  • L. Han
  • N. Fikadu
  • L. G. Thiam
  • A. Ba
  • R. Li
  • M. N. Pouye
  • K. Mangou
  • A. J. Moore
  • S. D. Sene
20 more authors
  • F. Diallo
  • E. M. Ngom
  • B. Sadio
  • A. Mbengue
  • C. Membi
  • B. Ngasala
  • T. Bazie
  • F. A. Some
  • N. Olson
  • S. D. Patel
  • L. Shapiro
  • S. Parikh
  • B. D. Foy
  • M. Cappello
  • I. Vigan-Womas
  • Z. Premji
  • R. K. Dabire
  • J.-B. Ouedraogo
  • Z. Sheng
  • A. K. Bei1
  1. 1Department of Epidemiology of Microbial Diseases, Yale School of Public Health, New Haven, CT, USA.
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Abstract

Transmission-blocking vaccines (TBVs) are a promising strategy to reduce malaria transmission by targeting parasite stages within the mosquito. However, parasite genetic diversity may limit vaccine efficacy. We used next-generation amplicon deep sequencing to identify non-synonymous single nucleotide polymorphisms (SNPs) in Pfs25 from 184 Plasmodium falciparum isolates from Senegal, Tanzania, Ghana, and Burkina Faso. Prioritized SNPs were introduced into P. falciparum via CRISPR-Cas9. For the G116C variant, gametocyte development was evaluated by microscopy and qPCR, and mosquito infectivity was assessed by SMFAs. We identified 26 SNPs, including 24 novel variants. Functional assays showed that the Pfs25 G116C mutation did not affect gametocyte development or exflagellation. SMFA showed no significant differences in oocyst prevalence or intensity between mutant and WT parasites. These findings highlight the importance of integrating genetic surveillance with functional validation to guide the development of effective transmission blocking interventions.

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Cite

BibTeXRIS
BibTeX
@misc{orfano2026identification,
  title = {{Identification of genetic variants in Pfs25 and functional evaluation in mosquito infection}},
  author = {A. Orfano and A. Cisse and Y. Guo and L. Han and N. Fikadu and L. G. Thiam and A. Ba and R. Li and M. N. Pouye and K. Mangou and A. J. Moore and S. D. Sene and F. Diallo and E. M. Ngom and B. Sadio and A. Mbengue and C. Membi and B. Ngasala and T. Bazie and F. A. Some and N. Olson and S. D. Patel and L. Shapiro and S. Parikh and B. D. Foy and M. Cappello and I. Vigan-Womas and Z. Premji and R. K. Dabire and J.-B. Ouedraogo and Z. Sheng and A. K. Bei},
  howpublished = {medRxiv},
  year = {2026},
  month = aug,
  doi = {10.64898/2026.08.25.26361130},
  url = {https://www.medrxiv.org/content/10.64898/2026.08.25.26361130v1},
  note = {CC BY-NC-ND},
}
RIS
TY  - UNPB
TI  - Identification of genetic variants in Pfs25 and functional evaluation in mosquito infection
AU  - A. Orfano
AU  - A. Cisse
AU  - Y. Guo
AU  - L. Han
AU  - N. Fikadu
AU  - L. G. Thiam
AU  - A. Ba
AU  - R. Li
AU  - M. N. Pouye
AU  - K. Mangou
AU  - A. J. Moore
AU  - S. D. Sene
AU  - F. Diallo
AU  - E. M. Ngom
AU  - B. Sadio
AU  - A. Mbengue
AU  - C. Membi
AU  - B. Ngasala
AU  - T. Bazie
AU  - F. A. Some
AU  - N. Olson
AU  - S. D. Patel
AU  - L. Shapiro
AU  - S. Parikh
AU  - B. D. Foy
AU  - M. Cappello
AU  - I. Vigan-Womas
AU  - Z. Premji
AU  - R. K. Dabire
AU  - J.-B. Ouedraogo
AU  - Z. Sheng
AU  - A. K. Bei
PY  - 2026
DA  - 2026/08/25
PB  - medRxiv
DO  - 10.64898/2026.08.25.26361130
UR  - https://www.medrxiv.org/content/10.64898/2026.08.25.26361130v1
AB  - Transmission-blocking vaccines (TBVs) are a promising strategy to reduce malaria transmission by targeting parasite stages within the mosquito. However, parasite genetic diversity may limit vaccine efficacy. We used next-generation amplicon deep sequencing to identify non-synonymous single nucleotide polymorphisms (SNPs) in Pfs25 from 184 Plasmodium falciparum isolates from Senegal, Tanzania, Ghana, and Burkina Faso. Prioritized SNPs were introduced into P. falciparum via CRISPR-Cas9. For the G116C variant, gametocyte development was evaluated by microscopy and qPCR, and mosquito infectivity was assessed by SMFAs. We identified 26 SNPs, including 24 novel variants. Functional assays showed that the Pfs25 G116C mutation did not affect gametocyte development or exflagellation. SMFA showed no significant differences in oocyst prevalence or intensity between mutant and WT parasites. These findings highlight the importance of integrating genetic surveillance with functional validation to guide the development of effective transmission blocking interventions.
ER  -