Abstract

ThegoalsofourR24projectincludeproducingnewrevertibleallelestoidentifygenes capableofpromotinghumanhealthandgeneratingnewmethodologiesforsite-directedsomatic mutagenesis,enhancinghomologousrecombinationandsingle-strandannealingmediated integrationofDNA.Itisanticipatedthatthesetechnologieswillgreatlyalleviatethenumberof animalsrequiredforgeneticstudiesinzebrafishandcanbeextendedtootheranimalspecies. Thesuccessofthistechnologydevelopmenthingesontheabilitytoscreenembryosandlarvae forfluorescentreportsofgeneexpressionandtransgenesis.InthisR24revision,weare proposingtoincreaseouropticalsectioningabilitiesandupdateexistingfluorescence microscopestomeettheincreaseddemandsonourimagingequipmentthatisacriticalpartof ourexperimentalstrategiesandthoseofothergroupsatIowaStateUniversity(ISU).These goalswillbeaccomplishedinthefollowingaims:
Aim1. Improveopticalsectioningcapabilities oflivingzebrafishembryosatIowaState.WeareproposingtopurchaseaZeissAxioImagerZ2 equippedwithanApotome2tomeettheincreasedopticalsectioningdemandsatISU.
Aim2. UpdateexistingmicroscopesusedinassociationwithourzebrafishfacilitieswithLEDlight sources.Together,theupdatesproposedabovewouldgreatlyincreaseourfluorescence screeningandimagingneedsofourcurrentlyfundedR24project,ourcolleaguesatISU,and futureinitiatives.

Public Health Relevance

ThegoalsofourR24projectaretotestnewrevertibleallelestoidentifygenescapableof promotinghumanhealth,todevelopnewmethodologiesforsite-directedsomaticmutagenesis, andtoenhancehomologousrecombinationandsingle-strandannealingmediatedintegrationof DNAinzebrafish.Usingfluorescencetoidentifyfounderswillgreatlyreducethenumberof zebrafishneededtoconductgenetargetingexperiments.Inthisrevision,werequestfundingfor state-of-theartmicroscopeequipmenttoenhanceourfluorescencescreeningandimaging capabilitiesandsuccessfullyachievethegoalsofourproject.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
Office of The Director, National Institutes of Health (OD)
Type
Resource-Related Research Projects (R24)
Project #
3R24OD020166-02S1
Application #
9324521
Study Section
Scientific and Technical Review Board on Biomedical and Behavioral Research Facilities (STOD)
Program Officer
Mccullough, Willie
Project Start
2016-06-01
Project End
2018-01-31
Budget Start
2017-05-01
Budget End
2018-01-31
Support Year
2
Fiscal Year
2017
Total Cost
Indirect Cost

  Institution

Name
Iowa State University
Department
Genetics
Type
Earth Sciences/Resources
DUNS #
005309844
City
Ames
State
IA
Country
United States
Zip Code
50011