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Cooperative Agreement N660011824505
Award Date
6/4/18
Completion Date
4/22/22
Dollars Obligated
$3.2M
Overview
Activity
4
Transactions
4
Subawards
2
Funding Federal Agency
Navy NAVSUP Engineering Logistics Office
Awarding Federal Agency
Naval Information Warfare Systems Command
Awardee
President And Fellows Of Harvard College (LN53LCFJFL45)
Federal Grant Program
12.910
Assistance Type
Cooperative Agreement
Place of Performance
Cambridge, MA 02138, USA
Description
Update #1
RAPID TESTS FOR SIGNATURES OF GENETIC ENGINEERING IN BIOLOGICAL SAMPLES
Posted 6/4/18, 12:00 AM
Mod #
Description
Reason For Modification
Federal Obligation
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Date
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Not listed
RAPID TESTS FOR SIGNATURES OF GENETIC ENGINEERING IN BIOLOGICAL SAMPLES - ADD SUBRECIPIENT, EXTEND TERM, UPDATE SOW AND MATRIX
$0
12/10/20
Not listed
P00002: MODIFICATION TO EXERCISE OPTION 1.
$970.9k
12/2/19
Not listed
INCREMENTAL FUNDING
$489.7k
8/7/18
Not listed
RAPID TESTS FOR SIGNATURES OF GENETIC ENGINEERING IN BIOLOGICAL SAMPLES
$1.7m
6/4/18
Grant Number
Description
Subgrantee
Prime Award
Dollars Obligated
(Click to sort descending)
Updated At
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16799105119225S
RAPID TESTS FOR SIGNATURES OF GENETIC ENGINEERING IN BIOLOGICAL SAMPLES OUR FELIX PROJECT AIMS TO DETECT, FROM CRUDE SAMPLES, POTENTIALLY HAZARDOUS DNA SEQUENCES THAT HAVE BEEN ENGINEERED OR ARE SIGNATURES OF DANGER. OUR APPROACH IS TO MODIFY BACTERIA THAT NATURAL TAKE UP DNA FROM THE ENVIRONMENT, SO THAT THE BACTERIA ARE HARD-WIRED TO RECOGNIZE THE POTENTIALLY HAZARDOUS SEQUENCES AND THEN PRODUCE A SIGNAL THAT CAN BE EASILY READ. THE ENTIRE DETECTION PROCESS WILL OCCUR IN AN HOUR AND WOULD ULTIMATELY BE CARRIED OUT BY A NON-SCIENTIST USING A SIMPLE KIT. OUR SYSTEM RESEMBLES A GEIGER COUNTER IN THAT A SINGLE MOLECULAR EVENT RAPIDLY GIVES RISE TO A MACROSCOPIC SIGNAL. TO ACHIEVE THIS, THE SYSTEM MUST BE POISED TO RELEASE STORED ENERGY THROUGH A CASCADE OF EVENTS THAT ARE HELD IN CHECK IN THE ABSENCE OF THE SIGNAL. TO ACCOMPLISH THIS, WE ARE ENGINEERING THE BACTERIUM BACILLUS SUBTILIS. B. SUBTILIS IS NATURALLY COMPETENT (I.E. IT NORMALLY TAKES UP DNA FROM THE EXTRACELLULAR ENVIRONMENT) AND IS GENETICALLY ENGINEERABLE, MAKING IT AN IDEAL HOST CELL TYPE FOR THIS BIOSENSOR. THIS B. SUBTILIS WILL BE ENGINEERED TO EXPRESS A CRISPR-BASED SYSTEM CONSISTING OF A DCAS9 PROTEIN FUSED TO A BACTERIAL PROTEIN KINASE AND TWO GUIDE RNAS THAT DIRECT THE DCAS9 TO BIND TO THE DNA SEQUENCE OF INTEREST. UPON BINDING, THE KINASE PROTEINS ARE BROUGHT INTO PROXIMITY, DIMERIZE TO BECOME ACTIVE, AND THEN PHOSPHORYLATE/ACTIVATE A TRANSCRIPTION FACTOR THAT THEN TURNS ON A REPORTER GENE. THE REPORTER GENE WILL BE CHOSEN TO PRODUCE A PRODUCT THAT IS EASILY DETECTABLE Â E.G, LIGHT, OR A COLORIMETRIC READOUT, ALLOWING A USER TO MAKE A QUICK JUDGEMENT ABOUT THE DNA CONTENTS OF A PARTICULAR SAMPLE. SUBCONTRACT FUNDS WILL BE USED TO PERFORM A PORTION OF PHASE 2 OF THIS PROJECT, INCLUDING FURTHER REFINEMENT OF THE B. SUBTILIS ENGINEERING, DETECTION AND PROOF OF CONCEPT VALIDATION OF AN EXPANDED NUMBER OF DNA SEQUENCES, CAPTURE OF DETECTED SEQUENCES, AS WELL AS REFINING, MULTIPLEXING, AND OPTIMIZING SYSTEM READOUTS AND OVERALL PERFORMANCE.
Northeastern University
Cooperative Agreement N660011824505
$0
2/18/22
1679915119225S
Rapid tests for signatures of genetic engineering in biological samples Our FELIX project aims to detect, from crude samples, potentially hazardous DNA sequences that have been engineered or are signatures of danger. Our approach is to modify bacteria that natural take up DNA from the environment, so that the bacteria are hard-wired to recognize the potentially hazardous sequences and then produce a signal that can be easily read. The entire detection process will occur in an hour and would ultimately be carried out by a non-scientist using a simple kit. Our system resembles a Geiger counter in that a single molecular event rapidly gives rise to a macroscopic signal. To achieve this, the system must be poised to release stored energy through a cascade of events that are held in check in the absence of the signal. To accomplish this, we are engineering the bacterium Bacillus subtilis. B. subtilis is naturally competent (i.e. it normally takes up DNA from the extracellular environment) and is genetically engineerable, making it an ideal host cell type for this biosensor. This B. subtilis will be engineered to express a CRISPR-based system consisting of a dCas9 protein fused to a bacterial protein kinase and two guide RNAs that direct the dCas9 to bind to the DNA sequence of interest. Upon binding, the kinase proteins are brought into proximity, dimerize to become active, and then phosphorylate/activate a transcription factor that then turns on a reporter gene. The reporter gene will be chosen to produce a product that is easily detectable â e.g, light, or a colorimetric readout, allowing a user to make a quick judgement about the DNA contents of a particular sample. Subcontract funds will be used to perform a portion of Phase 2 of this project, including further refinement of the B. subtilis engineering, detection and proof of concept validation of an expanded number of DNA sequences, capture of detected sequences, as well as refining, multiplexing, and optimizing system readouts and overall performance.
Northeastern University
Cooperative Agreement N660011824505
$452.8k
3/15/21