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Amplification-free detection of specific Leishmania species using CRISPR-Cas12

Award Number: F30AI191604
ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: FELLOWSHIP/SCHOLARSHIP/STUDENT LOANS
PERIOD OF PERFORMANCE START DATE: 05/01/2026
PERIOD OF PERFORMANCE END DATE: 04/30/2030

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Amplification-free detection of specific Leishmania species using CRISPR-Cas12 - PROJECT SUMMARY/ABSTRACT Leishmaniasis is a neglected tropical disease caused by over 20 protozoal species and transmitted by 70 species of sandflies, resulting in mucocutaneous, cutaneous, and visceral manifestations. Accurate species-level identification is essential for effective treatment and prognostication. Traditional microscopy lacks sensitivity and species specificity, whereas PCR, the most accurate current method, requires complex equipment and trained personnel in conventional laboratory settings. CRISPR-Cas direct detection technology, specifically CRISPR-Cas12, presents a promising alternative for species-specific diagnosis at the point of care. This technology detects DNA in vitro using a guide RNA that produces a fluorescent signal upon recognizing target DNA sequences, providing a faster and simpler alternative to PCR that could be used at the point of care. The objective of this research is to develop a CRISPR-based assay capable of speciating Leishmania, leveraging our expertise in CRISPR diagnostics for infectious diseases. This assay aims to enhance disease management by enabling accurate, rapid, and accessible species-level diagnosis. The project will achieve this by (1) identifying species-specific sequences for CRISPR-Cas targeting, and (2) developing and confirming Cas12 guides that can speciate Leishmania spp. DNA samples. These objectives will be accomplished through Cas12 guide design and selection, optimization of the CRISPR assay, and sample preparation to quantify parasite DNA spiked into blood. The anticipated outcomes include species-specific guides that be used for speciation, thereby informing treatment decisions. Additionally, the research will establish a pipeline for guide design applicable to further diagnostic research on Leishmaniasis and other neglected tropical diseases. The ultimate goal is to integrate this assay into a diagnostic platform suitable for low-resource settings, accelerating efforts to elucidate epidemiological patterns of leishmaniasis and guide treatment strategies aimed at disease elimination. These research objectives will be pursued in the context of a comprehensive research training program at the University of California, San Francisco, and Berkeley. This program encompasses structured technical and conceptual mentorship from the primary sponsor, Dr. Daniel Fletcher, an expert in diagnostics for neglected tropical diseases, and the co-sponsor, Dr. Dawn Wetzel, a physician-scientist expert in molecular parasitology. The applicant will receive additional training through structured preceptorships and regular advising sessions with physician-scientists specializing in the epidemiology and diagnostics of neglected tropical diseases, with a focus on pediatric care. This combination of experiences will provide invaluable training for an aspiring infectious disease physician-scientist.


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 = $43,097 )
2026	2026	REGENTS OF THE UNIVERSITY OF CALIFORNIA, SAN FRANCISCO, THE	1855 FOLSOM ST STE 425	SAN FRANCISCO	CA	94103	SAN FRANCISCO	USA	Allergy and Infectious Diseases Research	001	1	4/22/2026	NEW	$0
2026	2026	REGENTS OF THE UNIVERSITY OF CALIFORNIA, SAN FRANCISCO, THE	1855 FOLSOM ST STE 425	SAN FRANCISCO	CA	94103	SAN FRANCISCO	USA	Allergy and Infectious Diseases Research	000	1	4/3/2026	NEW	$43,097
														Subtotal = $43,097

Grand Total All Awards = $43,097

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Amplification-free detection of specific Leishmania species using CRISPR-Cas12

Award Number: F30AI191604

ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: FELLOWSHIP/SCHOLARSHIP/STUDENT LOANS

PERIOD OF PERFORMANCE START DATE: 05/01/2026

PERIOD OF PERFORMANCE END DATE: 04/30/2030

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