The Twin Spot Generator for differential Drosophila lineage analysis

  title={The Twin Spot Generator for differential Drosophila lineage analysis},
  author={Ruth Griffin and Anne Sustar and Marianne Bonvin and Richard Binari and Alberto Del Valle Rodr{\'i}guez and Chris Bakal and Amber M. Hohl and Jack R. Bateman and Christians Villalta and Elleard Heffern and Didier Grunwald and Claude Desplan and Gerold Schubiger and C.-ting Wu and Norbert Perrimon},
  journal={Nature methods},
  pages={600 - 602}
In Drosophila melanogaster, widely used mitotic recombination–based strategies generate mosaic flies with positive readout for only one daughter cell after division. To differentially label both daughter cells, we developed the twin spot generator (TSG) technique, which through mitotic recombination generates green and red twin spots that are detectable after the first cell division as single cells. We propose wide applications of TSG to lineage and genetic mosaic studies. 

Generating mosaics for lineage analysis in flies

  • Tzumin Lee
  • Biology
    Wiley interdisciplinary reviews. Developmental biology
  • 2014
Tracing neuronal lineages by using mosaics revolutionized mechanistic studies of neuronal diversification and differentiation, exemplifying the power of genetic mosaics in developmental biology.

MAGIC: Mosaic Analysis by gRNA-Induced Crossing-over

Eliminating the need to introduce recombinase-recognition sites in the genome, this simple and versatile system simplifies mosaic analysis in Drosophila and can be applied in any organism that is compatible with CRISPR/Cas9.

Power tools for gene expression and clonal analysis in Drosophila

A hands-on user guide to the techniques and approaches that have greatly increased resolution of genetic analysis in the fly, with a special focus on their application for lineage analysis.

Mosaic Analysis in Drosophila

How genetic mosaicism in Drosophila became an invaluable research tool that revolutionized developmental biology is discussed and methods ranging from simple “twin-spot” analysis to more sophisticated systems of multicolor labeling are covered.

New genetic tools for cell lineage analysis in Drosophila

Real-time lineage tracing in flies gets a boost with three techniques to specifically label a progenitor's daughter cells.

Versatile CRISPR/Cas9-mediated mosaic analysis by gRNA-induced crossing-over for unmodified genomes

Eliminating the need to introduce recombinase-recognition sites into the genome, this simple and versatile system simplifies mosaic analysis in Drosophila and can in principle be applied in any organism that is compatible with CRISPR/Cas9.

Drosophila twin spot clones reveal cell division dynamics in regenerating imaginal discs.

TIE-DYE: a combinatorial marking system to visualize and genetically manipulate clones during development in Drosophila melanogaster

The TIE-DYE method is developed that allows simultaneous lineage tracing of multiple cell populations as well as the genetic manipulation of a subset of these populations, allowing an assessment of the consequences of alterations in growth, patterning and cell-cell affinity.

Using MARCM to study Drosophila brain development.

The present protocol provides a detailed description of a standard MARCM analysis of brain development that includes generation of MARCM stocks and crosses, induction of clones, brain dissection at various stages of development, immunohistochemistry, and confocal microscopy, and can be modified for similar experiments involving mitotic clones.

Advanced Techniques for Cell Lineage Labelling in Drosophila

The ability to mark and genetically manipulate clonally related cells in live organisms is invaluable for investigating the mechanisms of tissue development, homeostasis and repair and creating a powerful arsenal of tools for the study of many diverse topics in tissue biology.



Site-specific recombination between homologous chromosomes in Drosophila.

A method of generating clones by inducing homologous mitotic recombination in Drosophila with a site-specific yeast recombinase is described, which allows for frequent mosaicism after mitotic exchange is induced at predefined sites in the genome.

The autosomal FLP-DFS technique for generating germline mosaics in Drosophila melanogaster.

This paper has built chromosomes that allow the use of the "FLP-DFS" technique to the autosomes, and describes the various steps involved in the development of this technique as well as the properties of the chromosomes utilized.

Genetic transformation of Drosophila with transposable element vectors.

A rosy transposon (ry1), constructed by inserting a chromosomal DNA fragment containing the wild-type rosy gene into a P transposable element, transformed germ line cells in 20 to 50 percent of the injected rosy mutant embryos indicating that the visible genetic defect in the host strain could be fully and permanently corrected by the transferred gene.

An optimized transgenesis system for Drosophila using germ-line-specific φC31 integrases

The efficiency, ease-of-use, and versatility obtained here with the φC31-based integration system represents an important advance in transgenesis and opens up the possibility of systematic, high-throughput screening of large cDNA sets and regulatory elements.

Transposition of cloned P elements into Drosophila germ line chromosomes.

Recombinant DNA carrying the 3-kilobase transposable element was injected into Drosophila embryos of a strain that lacked such elements, and observations provide the basis for efficient DNA-mediated gene transfer in Dosophila.

Site-Specific Transformation of Drosophila via ϕC31 Integrase-Mediated Cassette Exchange

A method for the precise targeting of transgenic constructs to predetermined genomic sites in Drosophila using the ϕC31 integrase system in conjunction with recombinase-mediated cassette exchange (RMCE) should greatly facilitate transgene analysis as well as permit new experimental designs.