Project Details
Description
PROJECT SUMMARY
This proposal seeks to develop a resource for the preservation of the fruit fly, Drosophila melanogaster. This
insect is a foundational model organism for biological research. Over a century of work, an enormous number
of fly strains harboring different mutant alleles or transgenic constructs have been generated. However, one
limitation of working with flies is that there is as yet no practical method for cryopreservation of Drosophila
strains. Conventional methods of vitrifying Drosophila were developed in the early 1990s and were never
widely adopted due to the difficulty in performing the protocols. This is a problem from a practical perspective
since all these strains need to be individually maintained in continuous culture at substantial cost and labor,
and also from a scientific perspective, since in the process of continuous culture mutations can accumulate
and contamination can occur, degrading the value of these resources for future experiments. A novel approach
for cryopreservation of Drosophila is proposed for this R24 resource center. Isolated embryonic nuclei, rather
than intact embryos, will be cryopreserved and then nuclear transplantation via microinjection will be used to
create clones derived from the cryopreserved nuclei. This approach avoids the issues associated with the
impermeability of embryonic membranes that have prevented the use of conventional cryopreservation
approaches that have been used with other organisms. Embryonic nuclei will be cryopreserved using a
naturally inspired approach. Diverse biological systems (plants, insects, etc.) survive dehydration, drought,
freezing temperatures and other stresses through the use of osmolytes. On an applied level, the proposed
investigation has the potential to transform preservation of Drosophila lines by 1) preserving subcellular
components (specifically nuclei) as opposed to embryos; and 2) automating much of the workflow. In the long-
term, the goal of this resource center is to develop a robust and scalable protocol for cryopreservation of
Drosophila, thus reducing the cost and improving the quality of long-term strain maintenance.
Status | Finished |
---|---|
Effective start/end date | 5/15/20 → 4/30/24 |
Funding
- NIH Office of the Director: $599,090.00
- NIH Office of the Director: $575,125.00
- NIH Office of the Director: $593,865.00
- NIH Office of the Director: $603,215.00
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