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MitoPac female killing systems in mosquitoes

Award Number: R21AI196522
ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 02/19/2026
PERIOD OF PERFORMANCE END DATE: 01/31/2028

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MitoPac female killing systems in mosquitoes - PROJECT SUMMARY Dengue, chikungunya, and Zika are diseases transmitted by Aedes aegypti. One strategy to reduce Ae. aegypti populations is to release non-biting male mosquitoes that pass female killing genes to their progeny. Over time and with many releases, this can lead to a significant suppression of the mosquito population, decreasing transmission of the pathogens they vector. Some female-killing technologies are available for this vector, but these technologies have some challenges that support the development of additional control technologies. To address this need, we aim to build new tools to generate female-killing technologies in Ae. aegypti. We focus on developing novel mitochondrial-targeted restriction endonucleases (mtREs) and CRISPR systems to kill females with flexible designs and components amenable for future development in a wide variety of mosquito vectors. As part of these efforts, we will explore switch-like cis-regulatory polycomb response elements (PREs) to modulate the transcriptional activity of the mtREs and Cas9. In Aim 1, we will selectively express the small mtRE, PacI, in female mosquitoes. Female-specific expression of PacI will be achieved by driving PacI expression with a female-specific promoter or by encoding a female-specific intron of the highly conserved doublesex (dsx) gene into the coding sequence of PacI. These expression systems will be further optimized by integrating the PRE upstream of the PacI promoter. In Aim 2, we will also use PREs to develop high precision heat shock-inducible and female-specific Cas9 expression systems. Heat shock promoters can still have activity at low temperatures, and Cas9 often has high activity, which can interfere with strain survival and the desired female-killing phenotype, so we aim to use PREs to minimize the issues. To achieve female specificity, the heat shock Cas9 will be designed to target female-specific essential genes, or the Cas9 will be engineered to encode a female-specific dsx intron in its coding sequence, ensuring Cas9 is only expressed in females. To optimize these Cas9 systems, they will be engineered first to target an easily screenable gene and then to target essential genes. The tools developed in Aims 1 and 2 will be evaluated in preliminary fitness and population cage studies, which will be used to prioritize their future development in Ae. aegypti and other mosquito vectors. These expression systems also have other applications for mosquito control and as a research tool to explore mosquito biology and genetics.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2026 ( Subtotal = $439,375 )
2026	2026	UNIVERSITY OF CALIFORNIA, SAN DIEGO	9500 GILMAN DR	LA JOLLA	CA	92093	SAN DIEGO	USA	Allergy and Infectious Diseases Research	000	1	2/19/2026	NEW	$439,375
														Subtotal = $439,375

Grand Total All Awards = $439,375

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MitoPac female killing systems in mosquitoes

Award Number: R21AI196522

ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

PERIOD OF PERFORMANCE START DATE: 02/19/2026

PERIOD OF PERFORMANCE END DATE: 01/31/2028

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