Research Advances and Challenges of Gene Drive Technology in Mosquito-Borne Disease Control

doi:10.12300/j.issn.1674-5817.2025.138

Abstract

Abstract:

Mosquito-borne diseases (such as malaria, dengue fever, Zika virus disease, and Chikungunya) pose major threats to global public health, while traditional control methods based on chemical pesticides face severe challenges including enhanced drug resistance in vector mosquitoes and environmental pollution. Genetic control strategies have become high-potential alternative solutions for mosquito control due to their species specificity and environmental friendliness. Gene drive technology uses gene editing tools such as clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated nuclease 9 (Cas9) to enable specific genes to efficiently spread in target mosquito populations through "super-Mendelian inheritance", offering a revolutionary strategy for the prevention and control of mosquito-borne diseases. This review systematically summarizes key advances, core challenges, and response strategies of gene drive technology in this field. Research advances: (1) In Anopheles malaria vectors, population suppression drives targeting sex determination genes or female reproductive genes can cause female sterility or skewed sex ratios to achieve population suppression. Population replacement gene drive strategies targeting host genes associated with Plasmodium infection or delivering anti-Plasmodium effector molecules in Anopheles can effectively block pathogen transmission. (2) In Aedes mosquito vectors of arboviruses, targeting female flight-essential genes achieves population suppression, and coupling of antiviral effector systems with drive elements is explored. Optimized split gene drive strategies demonstrate high cutting and recombination efficiency, and models predict safe and controllable spread of disease-resistance traits. (3) In Culex mosquitoes transmitting lymphatic filariasis, homology drive elements are integrated into two genes involved in the eye pigment synthesis pathway, allowing clear visualization of gene drive efficiency through eye color. Core Challenges: technological challenges include low homologous recombination repair efficiency, non-homologous end joining repair causing resistance allele generation, CRISPR/Cas9 off-target effects, and species adaptation differences. Ecological and safety challenges involve gene pool pollution caused by accidental spread of drive elements, potential ecological balance impacts, and long-term irreversible risks. Response strategies and prospects: employing multiplex guide RNA (gRNA) targeting strategies to enhance drive stability and combat potential resistance. Developing reversible designs such as synthetic resistance, reversal drives, and immunizing reversal drives as "genetic brakes". Establishing long-term ecological monitoring systems and mathematical modeling for risk assessment. Exploring "environmentally responsive drives" to enhance controllability. Future research should continuously optimize drive efficiency and specificity, deepen ecological risk evaluation, strengthen international cooperation, and promote ethical consensus and regulatory framework construction, with the aim of making gene drive technology a sustainable prevention and control strategy to address the global health challenge of mosquito-borne diseases under the premise of safety and controllability.

Key words: Gene drive, Mosquito-borne diseases, Population suppression/population replacement, CRISPR/Cas9

CLC Number:

Q95-33

YUN Jiaqi,MA Qin,WANG Guandong,et al. Research Advances and Challenges of Gene Drive Technology in Mosquito-Borne Disease Control[J]. Laboratory Animal and Comparative Medicine, 2025, 45(6): 773-783. DOI: 10.12300/j.issn.1674-5817.2025.138.

Figures/Tables 3

References 50

[1]	World Health Organization. Vector-borne diseases[Z/OL]. (2024-09-26)[2025-08-21] .
[2]	World Health Organization. Dengue[Z/OL]. (2025-08-21)[2025-08-21]. .
[3]	World Health Organization. New WHO guidelines for clinical management of arboviral diseases: dengue, chikungunya, Zika and yellow fever[Z/OL]. (2025-07-10)[2025-08-21]. .
[4]	KITTAYAPONG P, NINPHANOMCHAI S, LIMOHPASMANEE W, et al. Combined sterile insect technique and incompatible insect technique: the first proof-of-concept to suppress Aedes aegypti vector populations in semi-rural settings in Thailand[J]. PLoS Negl Trop Dis, 2019, 13(10):e0007771. DOI:10.1371/journal.pntd.0007771 .
[5]	HARRIS A F, MCKEMEY A R, NIMMO D, et al. Successful suppression of a field mosquito population by sustained release of engineered male mosquitoes[J]. Nat Biotechnol, 2012, 30(9):828-830. DOI:10.1038/nbt.2350 .
[6]	MCMENIMAN C J, LANE R V, CASS B N, et al. Stable introduction of a life-shortening Wolbachia infection into the mosquito Aedes aegypti [J]. Science, 2009, 323(5910):141-144. DOI:10.1126/science.1165326 .
[7]	PHUC H K, ANDREASEN M H, BURTON R S, et al. Late-acting dominant lethal genetic systems and mosquito control[J]. BMC Biol, 2007, 5:11. DOI:10.1186/1741-7007-5-11 .
[8]	HOFFMANN A A, MONTGOMERY B L, POPOVICI J, et al. Successful establishment of Wolbachia in Aedes populations to suppress dengue transmission[J]. Nature, 2011, 476(7361):454-457. DOI:10.1038/nature10356 .
[9]	AMUZU H E, TSYGANOV K, KOH C, et al. Wolbachia enhances insect-specific flavivirus infection in Aedes aegypti mosquitoes[J]. Ecol Evol, 2018, 8(11):5441-5454. DOI:10.1002/ece3.4066 .
[10]	ZÉLÉ F, NICOT A, BERTHOMIEU A, et al. Wolbachia increases susceptibility to Plasmodium infection in a natural system[J]. Proc Biol Sci, 2014, 281(1779):20132837. DOI:10.1098/rspb.2013.2837 .
[11]	BURT A. Site-specific selfish genes as tools for the control and genetic engineering of natural populations[J]. Proc Biol Sci, 2003, 270(1518):921-928. DOI:10.1098/rspb.2002.2319 .
[12]	AKBARI O S, BELLEN H J, BIER E, et al. BIOSAFETY. Safeguarding gene drive experiments in the laboratory[J]. Science, 2015, 349(6251):927-929. DOI:10.1126/science.aac7932 .
[13]	MARSHALL J M, HAY B A. Confinement of gene drive systems to local populations: a comparative analysis[J]. J Theor Biol, 2012, 294:153-171. DOI:10.1016/j.jtbi.2011.10.032 .
[14]	CHAMPER J, BUCHMAN A, AKBARI O S. Cheating evolution: engineering gene drives to manipulate the fate of wild populations[J]. Nat Rev Genet, 2016, 17(3):146-159. DOI:10.1038/nrg.2015.34 .
[15]	JAMES A A. Gene drive systems in mosquitoes: rules of the road[J]. Trends Parasitol, 2005, 21(2):64-67. DOI:10.1016/j.pt.2004.11.004 .
[16]	O'BROCHTA D A, ALFORD R T, PILITT K L, et al. piggyBac transposon remobilization and enhancer detection in Anopheles mosquitoes[J]. Proc Natl Acad Sci USA, 2011, 108(39):16339-16344. DOI:10.1073/pnas.1110628108 .
[17]	CONG L, RAN F A, COX D, et al. Multiplex genome engineering using CRISPR/Cas systems[J]. Science, 2013, 339(6121):819-823. DOI:10.1126/science.1231143 .
[18]	KYROU K, HAMMOND A M, GALIZI R, et al. A CRISPR-Cas9 gene drive targeting doublesex causes complete population suppression in caged Anopheles gambiae mosquitoes[J]. Nat Biotechnol, 2018, 36(11):1062-1066. DOI:10.1038/nbt.4245 .
[19]	HAMMOND A, GALIZI R, KYROU K, et al. A CRISPR-Cas9 gene drive system targeting female reproduction in the malaria mosquito vector Anopheles gambiae [J]. Nat Biotechnol, 2016, 34(1):78-83. DOI:10.1038/nbt.3439 .
[20]	DONG Y M, SIMÕES M L, MAROIS E, et al. CRISPR/Cas9-mediated gene knockout of Anopheles gambiae FREP1 suppresses malaria parasite infection[J]. PLoS Pathog, 2018, 14(3):e1006898. DOI:10.1371/journal.ppat.1006898 .
[21]	CARBALLAR-LEJARAZÚ R, OGAUGWU C, TUSHAR T, et al. Next-generation gene drive for population modification of the malaria vector mosquito, Anopheles gambiae [J]. Proc Natl Acad Sci USA, 2020, 117(37):22805-22814. DOI:10.1073/pnas.2010214117 .
[22]	GALIZI R, DOYLE L A, MENICHELLI M, et al. A synthetic sex ratio distortion system for the control of the human malaria mosquito[J]. Nat Commun, 2014, 5:3977. DOI:10.1038/ncomms4977 .
[23]	XU X J, CHEN J H, WANG Y, et al. Gene drive-based population suppression in the malaria vector Anopheles stephensi [J]. Nat Commun, 2025, 16(1):1007. DOI:10.1038/s41467-025-56290-2 .
[24]	LI Z Q, DONG Y M, YOU L, et al. Driving a protective allele of the mosquito FREP1 gene to combat malaria[J]. Nature, 2025, 645(8081):746-754. DOI:10.1038/s41586-025-09283-6 .
[25]	GREEN E I, JAOUEN E, KLUG D, et al. A population modification gene drive targeting both Saglin and Lipophorin impairs Plasmodium transmission in Anopheles mosquitoes[J]. eLife, 2023, 12:e93142. DOI:10.7554/eLife.93142 .
[26]	CARBALLAR-LEJARAZÚ R, DONG Y M, PHAM T B, et al. Dual effector population modification gene-drive strains of the African malaria mosquitoes, Anopheles gambiae and Anopheles coluzzii [J]. Proc Natl Acad Sci USA, 2023, 120(29):e2221118120. DOI:10.1073/pnas.2221118120 .
[27]	GANTZ V M, JASINSKIENE N, TATARENKOVA O, et al. Highly efficient Cas9-mediated gene drive for population modification of the malaria vector mosquito Anopheles stephensi [J]. Proc Natl Acad Sci USA, 2015, 112(49):E6736-E6743. DOI:10.1073/pnas.1521077112 .
[28]	MUÑOZ D, JIMENEZ A, MARINOTTI O, et al. The AeAct-4 gene is expressed in the developing flight muscles of female Aedes aegypti [J]. Insect Mol Biol, 2004, 13(5):563-568. DOI:10.1111/j.0962-1075.2004.00519.x .
[29]	O'LEARY S, ADELMAN Z N. CRISPR/Cas9 knockout of female-biased genes AeAct-4 or myo-fem in Ae. aegypti results in a flightless phenotype in female, but not male mosquitoes[J]. PLoS Negl Trop Dis, 2020, 14(12):e0008971. DOI:10.1371/journal.pntd.0008971 .
[30]	FRANZ A W E, SANCHEZ-VARGAS I, ADELMAN Z N, et al. Engineering RNA interference-based resistance to dengue virus type 2 in genetically modified Aedes aegypti [J]. Proc Natl Acad Sci USA, 2006, 103(11):4198-4203. DOI:10.1073/pnas.0600479103 .
[31]	WILLIAMS A E, SANCHEZ-VARGAS I, REID W R, et al. The antiviral small-interfering RNA pathway induces zika virus resistance in transgenic Aedes aegypti [J]. Viruses, 2020, 12(11):1231. DOI:10.3390/v12111231 .
[32]	BUCHMAN A, GAMEZ S, LI M, et al. Broad dengue neutralization in mosquitoes expressing an engineered antibody[J]. PLoS Pathog, 2020, 16(1):e1008103. DOI:10.1371/journal.ppat.1008103 .
[33]	REID W R, OLSON K E, FRANZ A W E. Current effector and gene-drive developments to engineer arbovirus-resistant Aedes aegypti (Diptera: Culicidae) for a sustainable population replacement strategy in the field[J]. J Med Entomol, 2021, 58(5):1987-1996. DOI:10.1093/jme/tjab030 .
[34]	LI M, YANG T, KANDUL N P, et al. Development of a confinable gene drive system in the human disease vector Aedes aegypti [J]. eLife, 2020, 9:e51701. DOI:10.7554/eLife.51701 .
[35]	FENG X C, LÓPEZ DEL AMO V, MAMELI E, et al. Optimized CRISPR tools and site-directed transgenesis towards gene drive development in Culex quinquefasciatus mosquitoes[J]. Nat Commun, 2021, 12(1):2960. DOI:10.1038/s41467-021-23239-0 .
[36]	FENG X C, KAMBIC L, NISHIMOTO J H K, et al. Evaluation of gene knockouts by CRISPR as potential targets for the genetic engineering of the mosquito Culex quinquefasciatus [J]. CRISPR J, 2021, 4(4):595-608. DOI:10.1089/crispr.2021.0028 .
[37]	HARVEY-SAMUEL T, FENG X C, OKAMOTO E M, et al. CRISPR-based gene drives generate super-Mendelian inheritance in the disease vector Culex quinquefasciatus [J]. Nat Commun, 2023, 14(1):7561. DOI:10.1038/s41467-023-41834-1 .
[38]	KANDUL N P, LIU J R, BUCHMAN A, et al. Assessment of a split homing based gene drive for efficient knockout of multiple genes[J]. G3 (Bethesda), 2020, 10(2):827-837. DOI:10.1534/g3.119.400985 .
[39]	CHAMPER J, REEVES R, OH S Y, et al. Novel CRISPR/Cas9 gene drive constructs reveal insights into mechanisms of resistance allele formation and drive efficiency in genetically diverse populations[J]. PLoS Genet, 2017, 13(7):e1006796. DOI:10.1371/journal.pgen.1006796 .
[40]	NOBLE C, ADLAM B, CHURCH G M, et al. Current CRISPR gene drive systems are likely to be highly invasive in wild populations[J]. eLife, 2018, 7:e33423. DOI:10.7554/eLife.33423 .
[41]	PROWSE T A A, CASSEY P, ROSS J V, et al. Dodging silver bullets: good CRISPR gene-drive design is critical for eradicating exotic vertebrates[J]. Proc Biol Sci, 2017, 284(1860):20170799. DOI: 10.1098/rspb.2017.0799 .
[42]	CHAMPER J, LIU J X, OH S Y, et al. Reducing resistance allele formation in CRISPR gene drive[J]. Proc Natl Acad Sci USA, 2018, 115(21):5522-5527. DOI:10.1073/pnas.1720354115 .
[43]	VELLA M R, GUNNING C E, LLOYD A L, et al. Evaluating strategies for reversing CRISPR-Cas9 gene drives[J]. Sci Rep, 2017, 7(1):11038. DOI: 10.1038/s41598-017-10633-2 .
[44]	ESVELT K M, SMIDLER A L, CATTERUCCIA F, et al. Concerning RNA-guided gene drives for the alteration of wild populations[J]. eLife, 2014, 3:e03401. DOI: 10.7554/eLife.03401 .
[45]	WU B, LUO L Q, GAO X J. Cas9-triggered chain ablation of cas9 as a gene drive brake[J]. Nat Biotechnol, 2016, 34(2):137-138. DOI:10.1038/nbt.3444 .
[46]	ADOLFI A, GANTZ V M, JASINSKIENE N, et al. Efficient population modification gene-drive rescue system in the malaria mosquito Anopheles stephensi [J]. Nat Commun, 2020, 11(1):5553. DOI: 10.1038/s41467-020-19426-0 .
[47]	SÁNCHEZ C H M, WU S L, BENNETT J B, et al. MGDrivE: a modular simulation framework for the spread of gene drives through spatially explicit mosquito populations[J]. Meth Ecol Evol, 2020, 11(2):229-239. DOI:10.1111/2041-210X.13318 .
[48]	NORTH A R, BURT A, GODFRAY H C J. Modelling the suppression of a malaria vector using a CRISPR-Cas9 gene drive to reduce female fertility[J]. BMC Biol, 2020, 18(1):98. DOI: 10.1186/s12915-020-00834-z .
[49]	OBERHOFER G, IVY T, HAY B A. Gene drive that results in addiction to a temperature-sensitive version of an essential gene triggers population collapse in Drosophila [J]. Proc Natl Acad Sci USA, 2021, 118(49):e2107413118. DOI: 10.1073/pnas.2107413118 .
[50]	WANG G D, VEGA-RODRÍGUEZ J, DIABATE A, et al. Clock genes and environmental cues coordinate Anopheles pheromone synthesis, swarming, and mating[J]. Science, 2021, 371(6527):411-415. DOI:10.1126/science.abd4359 .

蚊虫品种 Mosquito species	靶标 Target	基因驱动类型 Gene drive type	参考文献 Reference
冈比亚按蚊 Anopheles gambiae	ribosomal gene	减数分裂介导的基因驱动	Galizi R, et al. (2014)^[22]
	doublesex	CRISPR/Cas9介导的基因驱动 CRISPR/Cas9介导的基因驱动 CRISPR/Cas9介导的基因驱动 CRISPR/Cas9介导的基因驱动 CRISPR/Cas9介导的基因驱动	Kyrou K, et al. (2018)^[18]
	female-sterility gene		Hammond A, et al. (2016)^[19]
	fibrinogen-related protein 1		Dong Y, et al. (2018)^[20]
	cardinal		Carballar-lejarazu R, et al. (2020)^[21]
	single chain variable fragment		Carballar-lejarazu R, et al. (2023)^[26]
斯氏按蚊 Anopheles stephensi	doublesex	CRISPR/Cas9介导的基因驱动 CRISPR/Cas9介导的基因驱动	Xu X, et al. (2025)^[23]
斯氏按蚊 Anopheles stephensi	kynurenine hydroxylase	CRISPR/Cas9介导的基因驱动 CRISPR/Cas9介导的基因驱动	Gantz V M, et al. (2015)^[27]
科卢兹按蚊 Anopheles coluzzii	Saglin	CRISPR/Cas9介导的基因驱动	Green E I, et al. (2023)^[25]
致倦库蚊 Culex quinquefasciatus	cardinal, white, kynurenine 3-monooxygenase	CRISPR/Cas9介导的基因驱动CRISPR/Cas9介导的基因驱动	Feng X C, et al. (2021)^[36]
埃及伊蚊 Aedes aegypti	AeAct-4, myo-fem	CRISPR/Cas9介导的基因驱动CRISPR/Cas9介导的基因驱动	O'Leary S, et al. (2020)^[29]
	white	CRISPR/Cas9介导的基因驱动CRISPR/Cas9介导的基因驱动	Li M, et al. (2020)^[34]
	inverted repeat RNA	归巢核酸内切酶介导的基因驱动	Franz A W, et al. (2006)^[30] Williams A E, et al. (2020)^[31]
	single chain variable fragment	转座子介导的基因驱动	Buchman A, et al. (2020)^[32]

蚊虫品种 Mosquito species	靶标 Target	基因驱动类型 Gene drive type	参考文献 Reference
冈比亚按蚊 Anopheles gambiae	ribosomal gene	减数分裂介导的基因驱动	Galizi R, et al. (2014)^[22]
	doublesex	CRISPR/Cas9介导的基因驱动 CRISPR/Cas9介导的基因驱动 CRISPR/Cas9介导的基因驱动 CRISPR/Cas9介导的基因驱动 CRISPR/Cas9介导的基因驱动	Kyrou K, et al. (2018)^[18]
	female-sterility gene		Hammond A, et al. (2016)^[19]
	fibrinogen-related protein 1		Dong Y, et al. (2018)^[20]
	cardinal		Carballar-lejarazu R, et al. (2020)^[21]
	single chain variable fragment		Carballar-lejarazu R, et al. (2023)^[26]
斯氏按蚊 Anopheles stephensi	doublesex	CRISPR/Cas9介导的基因驱动 CRISPR/Cas9介导的基因驱动	Xu X, et al. (2025)^[23]
斯氏按蚊 Anopheles stephensi	kynurenine hydroxylase	CRISPR/Cas9介导的基因驱动 CRISPR/Cas9介导的基因驱动	Gantz V M, et al. (2015)^[27]
科卢兹按蚊 Anopheles coluzzii	Saglin	CRISPR/Cas9介导的基因驱动	Green E I, et al. (2023)^[25]
致倦库蚊 Culex quinquefasciatus	cardinal, white, kynurenine 3-monooxygenase	CRISPR/Cas9介导的基因驱动CRISPR/Cas9介导的基因驱动	Feng X C, et al. (2021)^[36]
埃及伊蚊 Aedes aegypti	AeAct-4, myo-fem	CRISPR/Cas9介导的基因驱动CRISPR/Cas9介导的基因驱动	O'Leary S, et al. (2020)^[29]
	white	CRISPR/Cas9介导的基因驱动CRISPR/Cas9介导的基因驱动	Li M, et al. (2020)^[34]
	inverted repeat RNA	归巢核酸内切酶介导的基因驱动	Franz A W, et al. (2006)^[30] Williams A E, et al. (2020)^[31]
	single chain variable fragment	转座子介导的基因驱动	Buchman A, et al. (2020)^[32]