Flip-out Gal4
Flip-out Gal4 - Related Videos
13:34
Gene Trapping Using Gal4 in Zebrafish
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Cited by 19 •
2013
This protocol describes the method of gene trap insertional mutagenesis using Gal4-VP16 as the primary reporter and GFP/RFP as secondary reporters in zebrafish. Approximately one in ten high-expressing F0 fish yield gene trap progeny co-expressing GFP and RFP. The screening procedure can be readily scaled to adapt to the size of the laboratory performing the insertional mutagenesis screen.
05:25
Patterning of Embryonic Stem Cells Using the Bio Flip Chip
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Cited by 2 •
2007
We demonstrate a simple method for placing cells at desired locations on a substrate. This method patterns cells by flipping a silicone chip containing microwells filled with cells onto the substrate. This method provides a new way to modulate diffusible and juxtacrine signaling between cells.
08:21
Laser-induced Forward Transfer for Flip-chip Packaging of Single Dies
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Cited by 5 •
2015
We demonstrate the use of the Laser-induced Forward Transfer (LIFT) technique for flip-chip assembly of optoelectronic components. This approach provides a simple, cost-effective, low-temperature, fast and flexible solution for fine-pitch bumping and bonding on chip-scale for achieving high-density circuits for optoelectronic applications.
06:29
Using the GAL4-UAS System for Functional Genetics in Anopheles gambiae
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Cited by 11 •
2021
The bipartite GAL4-UAS system is a versatile tool for modification of gene expression in a controlled spatiotemporal manner which permits functional genetic analysis in Anopheles gambiae. The procedures described for using this system are a semi-standardized cloning strategy, sexing and screening of pupae for fluorescent protein markers and embryo fixation.
09:18
Photobleaching Assays (FRAP & FLIP) to Measure Chromatin Protein Dynamics in Living Embryonic Stem Cells
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Cited by 26 •
2011
We describe photobleaching methods including Fluorescence Recovery After Photobleaching (FRAP) and Fluorescence Loss In Photobleaching (FLIP) to monitor chromatin protein dynamics in embryonic stem (ES) cells. Chromatin protein dynamics, which is considered to be one of the means to study chromatin plasticity, is enhanced in pluripotent cells.
FAQs
FLP recombinase acts as the molecular switch in this system. Following heat-shock induction, FLP catalyzes recombination between the FRT sites surrounding a transcriptional stop cassette. Removing that cassette permits Gal4 expression in the recombined cells, which can then activate linked UAS reporters or effectors. This design converts a transient induction event into clone-restricted genetic activity.
Recombination occurs in selected cells after induction, so only those cells lose the stop cassette and express Gal4. Their descendants retain the activated state, producing defined clones within otherwise nonactivated tissue. This mosaic organization allows researchers to compare manipulated and neighboring unmanipulated cells in the same developing or tumor-bearing tissue.
Gal4 provides a common transcriptional driver, while UAS-linked components determine what the activated clone reveals or does. A reporter makes the clone visible for lineage tracing and clonal analysis; an effector enables targeted gene perturbation. Using these interchangeable outputs lets investigators connect a defined cellular population with changes in proliferation, survival, invasion, or tissue organization.
The workflow begins with heat-shock induction to activate FLP recombinase. FLP then recombines the paired FRT sites and removes the transcriptional stop cassette in selected cells. Gal4 expression subsequently activates the chosen UAS-linked reporter or cancer-related effector. Researchers examine the resulting clones in developing or tumor-bearing tissues to assess cellular behavior and tissue-level effects.
This approach is useful when the question concerns the behavior of defined cell populations within a larger tissue context. Clonal activation permits lineage tracing, direct clonal analysis, and targeted perturbation while preserving surrounding cells for comparison. In cancer research, that arrangement helps associate oncogenic changes with local effects on proliferation, survival, invasion, and tissue organization.
By activating reporters or effectors in selected clones, the system can reveal how oncogenic mutations influence tumor-related behaviors. Analyses may focus on altered cell proliferation, survival, invasion, or organization within developing or tumor-bearing tissues. These observations help connect genetic perturbations to tumor initiation and progression rather than viewing those processes only at the level of whole tissues.
Flip-out Gal4 provides a flexible platform for targeted gene perturbation in defined cellular clones. Researchers can observe how changing a candidate factor affects clone behavior and tissue organization, then use the resulting patterns to evaluate its contribution to tumor initiation or progression. The same framework can support studies of candidate mechanisms and potential interventions in Drosophila cancer models.