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New toolkit to visualize RNAs in living cells

Award Number: R44MH096366
ORGANIZATION: NATIONAL INSTITUTE OF MENTAL HEALTH
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 06/01/2012
PERIOD OF PERFORMANCE END DATE: 06/30/2018

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New toolkit to visualize RNAs in living cells - DESCRIPTION (provided by applicant): Over the past 10 years it has become increasingly apparent that there is exceptional complexity in the RNA population in cells. This diverse RNA pool includes microRNAs, Piwi-interacting RNAs, termini- associated RNAs and other noncoding RNAs. In many cases, alterations in these RNAs, or proteins that bind to these RNAs, have been linked to a wide range of medical disorders. A major challenge of molecular biology is to determine the function of these fascinating and novel RNA species. In order to understand how these RNAs function in cells, it is highly useful to image the localization and intracellular movements of these RNAs in living cells under a variety of experimental stimuli. We have developed a novel genetically encodable system to fluorescently tag RNAs in cells. This system utilizes an RNA sequence element, termed Spinach, which is appended to an RNA of interest, and "switches on" the fluorescence of an otherwise non-florescent dye. This dye is based on the structure of the fluorophore in GFP, making Spinach an RNA mimic of GFP. In the Phase I portion of this project we markedly increased the brightness of Spinach in cells, and expanded the use of Spinach to novel noncoding RNAs. We also developed novel expression plasmids for expressing Spinach- tagged RNAs. In this Phase II proposal, we will develop novel concatamerization strategies to label RNAs with tandem repeats of Spinach, thereby increasing the sensitivity of this labeling approach. We also will tag a set of major noncoding RNAs that label cellular structures, in order to provide a suite of plasmids expressing pre-labeled RNAs that are ready to use for researchers. We also will expand the spectral imaging capacity of this approach by developing new fluorophores for Spinach, and new RNA-fluorophore complexes, which will provide the spectral versatility in biologic imaging that is needed by the research community. Based on the past history of successful commercialization of GFP expression systems, we expect that this expression system will have high commercial potential and will accelerate the pace of RNA research.


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: 2019 ( Subtotal = $0 )
2019	2016	LUCERNA INC.	760 PARKSIDE AVE STE 311	BROOKLYN	NY	11226	KINGS	USA	Mental Health Research Grants	000	4	11/29/2018	NON-COMPETING CONTINUATION	$0
														Subtotal = $0

Issue Date FY: 2016 ( Subtotal = $237,583 )
2016	2016	LUCERNA INC	POST OFFICE BOX 1342	NEW YORK	NY	10021	NEW YORK	USA	Mental Health Research Grants	000	4	7/12/2016	NON-COMPETING CONTINUATION	$237,583
														Subtotal = $237,583

Issue Date FY: 2015 ( Subtotal = $512,658 )
2015	2015	LUCERNA INC	POST OFFICE BOX 1342	NEW YORK	NY	10021	NEW YORK	USA	Mental Health Research Grants	001	3	8/5/2015	NON-COMPETING CONTINUATION	$507,522
2015	2014	LUCERNA INC	POST OFFICE BOX 1342	NEW YORK	NY	10021	NEW YORK	USA	Mental Health Research Grants	000	2	7/21/2015	COMPETING CONTINUATION	$5,136
														Subtotal = $512,658

Issue Date FY: 2014 ( Subtotal = $498,492 )
2014	2014	LUCERNA INC	POST OFFICE BOX 1342	NEW YORK	NY	10021	NEW YORK	USA	Mental Health Research Grants	000	2	6/24/2014	COMPETING CONTINUATION	$498,492
														Subtotal = $498,492

Grand Total All Awards = $1,248,733

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New toolkit to visualize RNAs in living cells

Award Number: R44MH096366

ORGANIZATION: NATIONAL INSTITUTE OF MENTAL HEALTH

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

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

PERIOD OF PERFORMANCE START DATE: 06/01/2012

PERIOD OF PERFORMANCE END DATE: 06/30/2018

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