6) One of your colleagues is brainstorming on how to use the Flp-FRT system to s

Question

6) One of your colleagues is brainstorming on how to use the Flp-FRT system to selectively remove gene X from serotonergic cells in adult flies. She wants to know how the absence of this gene will affect serotonergic neurotransmission in the fly brain. She has drawn out two models on how to build the transgenic flies and would like your opinion (see Figure 1). Here you see the use ofa two component system to delete certain exons from gene X (shown in grey). After a quick glance you realize that one of these models will probably not work to remove protein X (as a product of gene X) in adult fly brains. In your estimation which transgenic fly is flawed? What are two things that are wrong with the system as designed in that transgenic fly? Along those lines, how is the other transgenic fly better designed? Last, how could you further improve that transgenic fly? What is your rationale behind the improvement?

Explanation / Answer

Transgenic fly 1 is flawed. This is because , for a FLP -FRT system a heat shock promoter is needed to induce site specific recombination. In the FLP -FRT system a holliday intermediate is created and later recombiantion takes place. In the transgenic fly 1 , the FRT region ie flippase target region is present between exon 2 to exon 4 which is unable to create a holliday junction model. This is because the exon 4 is the last region before STOP codon and the holliday model can be created only in between the genome and not at the ends. So transgenic fly 1 is flawed . In transgenic fly 2 , heat shock gene is present and also the FRT sequence is in the middle . So holliday intermediate model can be created and the recombination can takes place. The transgenic fly 2 can be further modified and improved by incorporating a gene for GFP protein. This transgenic fly with transgenic protein gene X can be viewed under fluroscence microscopy when produced.

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