Flp-frt System
Flp-frt System - Related Videos
09:33
The Tomato/GFP-FLP/FRT Method for Live Imaging of Mosaic Adult Drosophila Photoreceptor Cells
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Cited by 25 •
2013
The Tomato/GFP-FLP/FRT method involves visualizing mosaic photoreceptor cells in living Drosophila. It can be used to follow individual photoreceptor cell fates in the retina for days or weeks. This method is ideal for studies of retinal degeneration and neurodegenerative diseases or photoreceptor cell development.
12:13
TransFLP — A Method to Genetically Modify Vibrio cholerae Based on Natural Transformation and FLP-recombination
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Cited by 31 •
2012
A quick method to modify the genome of V. cholerae is described. These modifications include the deletion of single genes, gene clusters and genomic islands as well as the integration of short sequences (e.g. promoter elements or affinity-tag sequences). The method is based on the natural transformation and FLP-recombination.
08:30
Application of MultiColor FlpOut Technique to Study High Resolution Single Cell Morphologies and Cell Interactions of Glia in Drosophila
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Cited by 4 •
2017
Cells display different morphologies and establish a variety of interactions with their neighbors. This protocol describes how to reveal the morphology of single cells and to investigate cell-cell interaction by using the well-established Gal4/UAS expression system.
10:31
The Drosophila Imaginal Disc Tumor Model: Visualization and Quantification of Gene Expression and Tumor Invasiveness Using Genetic Mosaics
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Cited by 10 •
2016
This protocol demonstrates how to generate fluorescently marked, genetically defined clonal tumors in the Drosophila eye/antennal imaginal discs (EAD). It describes how to dissect the EAD and brain from the third instar larvae and how to process them to visualize and quantify gene expression changes and tumor invasiveness.
05:48
A Behavioral Assay for Mechanosensation of MARCM-based Clones in Drosophila melanogaster
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Cited by 1 •
2015
In order to identify novel mutations affecting mechanosensation, we designed an assay that measures the behavioral response to tactile stimulation of fly bristles in mutant clones generated by the MARCM method. The combination of techniques allows for the identification of mechanosensitive mutations that would otherwise be lethal.
FAQs
FRT-site orientation determines whether the intervening DNA is excised or inverted. When paired sites run in parallel, Flp recombinase can remove the sequence between them. When the sites face opposite directions, the intervening segment can be inverted instead. Designing site orientation therefore allows researchers to select the type of genetic rearrangement required for a particular experiment.
Flp recombinase supplies the enzymatic activity, while FRT sites provide the defined DNA locations recognized by that enzyme. Their partnership restricts rearrangement to DNA segments positioned between matching sites, rather than producing an unspecified change across the genome. This division between enzyme and recognition sequence gives the method its precise and programmable character in molecular genetics.
Predictable rearrangements make it possible to plan genetic changes around the location and orientation of FRT sites. Researchers can remove a selectable marker, alter the arrangement of a transgene, or create a conditional modification without treating the outcome as random. This control is especially valuable when experiments require reproducible DNA organization or regulated gene manipulation.
The target sequence must be placed between a pair of FRT sites, with their relative orientation selected according to the desired outcome. Parallel placement supports excision, whereas opposite orientation supports inversion. This design principle lets investigators connect the physical arrangement of sites in an engineered DNA construct with the rearrangement they intend to study.
A conceptual workflow begins by designing DNA that contains the sequence of interest and an appropriate pair of FRT sites, then providing Flp recombinase in the relevant cells or organism. The resulting DNA rearrangement is examined in relation to the intended excision or inversion. The same logic can support marker removal, conditional modification, or transgene reorganization.
Researchers apply the system when a study needs controlled changes to engineered DNA in cells or organisms. Supported uses include removing selectable marker genes, creating conditional genetic modifications, and controlling transgene organization. In biochemistry and transgenic model development, these applications help connect defined DNA rearrangements with regulated gene manipulation and the analysis of resulting experimental outcomes.