Friday, August 29, 2025 8/29/2025

Characterizing mechanisms of immune protection by the bacterial type II-A CRISPR systems

Award Number: R03AI175981
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: 12/12/2023
PERIOD OF PERFORMANCE END DATE: 10/31/2025

Group Awards By:

View Award Description

Characterizing mechanisms of immune protection by the bacterial type II-A CRISPR systems - Project Summary/Abstract Bacteria employ several distinct defense strategies for protection from phage infection. CRISPR-Cas is one such mechanism that provides an adaptive immune protection against recurring phage infections based on genetic memory retained from past infections. The genetic memory is retained through a process called “adaptation” where short pieces of the intruder DNA (called prespacers) are selected following certain rules, and site-specifically inserted into the CRISPR locus as a “spacer”. The spacer is transcribed into guide-RNAs that are essential for sequence-specific targeting and inactivation of foreign genetic material. Adaptation in type II- A CRISPR systems is unique compared to other CRISPR systems due to the inherent capabilities of Cas1 and Cas2 proteins to catalyze the site-specific prespacer insertion without assistance from cellular factors to maintain fidelity during insertion. This property offers unique promises for type II-A Cas1-Cas2-based biotechnological and biomedical applications. Previous work from the PI’s group has identified distinct subgroups of type II-A CRISPR systems based on conserved DNA motifs present at the site of insertion, which is conserved across many bacterial genera. Accompanying work had biochemically established unique differences in the mechanisms of prespacer insertion by the different subgroups. The proposed research aims to derive molecular mechanisms of the differences in DNA requirements and efficiencies of prespacer insertion between the different type II-A subgroups. To derive structural and conformational differences between these subgroups, hydrogen deuterium exchange mass spectrometry (HDX-MS) and molecular dynamics (MD) simulations will be performed, followed by biochemical validation of the results from HDX-MS and MD simulations. To assess the contribution of individual protein and nucleic acid complexes in site-specific DNA insertion, QM/MM (quantum mechanics/molecular mechanics) calculations will be employed. Based on the free energy contributions identified, amino acids and DNA nucleotides will be mutated to change integration efficiency with the end goal of engineering new Cas1-Cas2 variants which has developed specificity to a newly designed DNA sequence. Since Cas1 and Cas2 based prespacer insertion is vital to the functioning of all CRISPR systems, the lessons learned can be translated to other CRISPR systems. Applications include programmed DNA insertions, enhancing phage therapy to fight drug-resistant bacteria, and to develop bacteria that can fight off phage infections for the fermentation industry.


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: 2025 ( Subtotal = $70,990 )
2025	2025	UNIVERSITY OF OKLAHOMA	660 PARRINGTON OVAL RM 301	NORMAN	OK	73019	CLEVELAND	USA	Allergy and Infectious Diseases Research	000	2	1/3/2025	NON-COMPETING CONTINUATION	$70,990
														Subtotal = $70,990

Issue Date FY: 2024 ( Subtotal = $70,990 )
2024	2024	UNIVERSITY OF OKLAHOMA	660 PARRINGTON OVAL RM 301	NORMAN	OK	73019	CLEVELAND	USA	Allergy and Infectious Diseases Research	001	1	5/29/2024	NEW	$7,099
2024	2024	UNIVERSITY OF OKLAHOMA	660 PARRINGTON OVAL RM 301	NORMAN	OK	73019	CLEVELAND	USA	Allergy and Infectious Diseases Research	000	1	12/12/2023	NEW	$63,891
														Subtotal = $70,990

Grand Total All Awards = $141,980

Top

All Categories

About

Search

Reports

Data Submission

Award Information

Characterizing mechanisms of immune protection by the bacterial type II-A CRISPR systems

Award Number: R03AI175981

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: 12/12/2023

PERIOD OF PERFORMANCE END DATE: 10/31/2025

Federal Websites

Department of Health & Human Services

HHS Operating Divisions

HHS Staff Divisions

Download A Document Viewer