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View full policyAn engineered protein tag resource for live imaging and functional studies in zebrafish
About This Grant
Project Summary Protein studies rely on highly specific antibodies that can function across a range of cell biological and biochemical experimental techniques. The zebrafish is an outstanding vertebrate genetic model organism for the study of development, physiology, behavior, and disease etiology. Yet, sufficient quality antibodies are not available for the majority of the zebrafish proteome. One objective of this application is to engineer a resource of multifunctional epitope-tagged zebrafish lines of high interest to the community and relevant to human disease. We have developed a software package to scan protein sequences for optimal epitope tag integration sites based on conservation of protein sequence and structural features, as well as machine-learning based protein structure prediction, where insertion of an epitope is most optimal to preserve fully protein function. We have tested the functionality of the program on two proteins identifying internal epitope tagging sites as optimal for epitope tagging. Consistent with the computational prediction, we experimentally determined that internal epitope tagged proteins rescue loss-of-function phenotypes similarly to wild type mRNA, while N- and/or Cterminally tagged proteins do not. In Specific Aim 1, we will continue to develop and empirically validate our user-friendly software package, and make our software freely available as a web server to researchers worldwide. Furthermore, we will poll the zebrafish community to identify an additional 60 genes of high interest and important to human health and disease and then generate epitope-tagged zebrafish lines of these genes. In Specific Aim 2, we will test the recenUy developed ALFA epitope tag as a method to investigate protein function in zebrafish. The ALFA-tag is recognized by a camel id antibody NbALFA, with the potential to bring any effector protein fused to NbALFA to the immediate proximity of the ALFA-tagged protein. We will develop a resource of transgenic NbALFA fusion resources for live visualization (GFP), degradation (Degron), proximity labeling (TurbolD} and identification of transcription factor target sites (DamlD}. As a result, a single genome edit (integration of the ALFA-tag) will enable comprehensive analysis of protein function.
Grant Summary
An engineered protein tag resource for live imaging and functional studies in zebrafish is a OD - NIH Office of the Director grant providing up to $812K for university, nonprofit, healthcare org. Applications are due 2030-03-31 (open). Check eligibility and apply with FindGrants.
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Eligibility
How to Apply
Up to $812K
2030-03-31
- 1Confirm your organization is eligible for An engineered protein tag resource for live imaging and functional studies in zebrafish from OD - NIH Office of the Director, checking organization type, location, and any population or project requirements.
- 2Gather the required documents and information, including your organization details, project plan, and budget figures.
- 3Draft your application narrative and budget addressing the funder's priorities and review criteria. FindGrants can draft each section for you to review and edit.
- 4Review every section against the requirements checklist, then export a submission-ready application pack and submit it to OD - NIH Office of the Director before the deadline.
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An engineered protein tag resource for live imaging and functional studies in zebrafish: Frequently Asked Questions
Who is eligible for the An engineered protein tag resource for live imaging and functional studies in zebrafish?
An engineered protein tag resource for live imaging and functional studies in zebrafish is offered by OD - NIH Office of the Director and is generally open to university, nonprofit, healthcare org. It is open to organizations nationwide unless the funder specifies otherwise. Review the specific eligibility terms before applying, since funders set their own requirements around organization type, location, and the population or project being served.
How much funding does the An engineered protein tag resource for live imaging and functional studies in zebrafish provide?
An engineered protein tag resource for live imaging and functional studies in zebrafish provides up to $812K per award from OD - NIH Office of the Director. Actual award sizes depend on the scope of your project, available program funds, and the number of applicants, so build a budget that reflects realistic, allowable costs rather than the maximum figure.
When is the An engineered protein tag resource for live imaging and functional studies in zebrafish deadline?
Applications for An engineered protein tag resource for live imaging and functional studies in zebrafish are due 2030-03-31 (open). Because deadlines can change, verify the date with the funder, OD - NIH Office of the Director, and give yourself enough time to prepare a complete, competitive application before the close date.
How do you apply for the An engineered protein tag resource for live imaging and functional studies in zebrafish?
To apply for An engineered protein tag resource for live imaging and functional studies in zebrafish, confirm your eligibility, gather the required documents, and prepare a narrative and budget that address the funder's priorities. FindGrants guides you step by step and can draft each section, then exports a submission-ready application pack for this grant from OD - NIH Office of the Director.