Related Experiment Video
Updated: Apr 22, 2026
10:01
An Efficient Strategy for Generating Tissue-specific Binary Transcription Systems in Drosophila by Genome Editing
Published on: September 19, 2018
8.5K
A versatile two-step CRISPR- and RMCE-based strategy for efficient genome engineering in Drosophila.
Xu Zhang1, Wouter H Koolhaas1, Frank Schnorrer2
1Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.
G3 (Bethesda, Md.)
|October 18, 2014
Summary
We developed an efficient CRISPR-RMCE strategy for flexible Drosophila melanogaster genome engineering. This two-step method allows rapid modification of multiple genomic locations, making gene targeting accessible to all fly labs.
Area of Science:
- Genetics and Genomics
- Molecular Biology
- Developmental Biology
Background:
- CRISPR/Cas gene editing offers powerful genome engineering capabilities for model organisms.
- CRISPR applications in Drosophila melanogaster are limited by variable efficiency and scope.
- Existing methods question the universal applicability of CRISPR for Drosophila gene targeting.
Purpose of the Study:
- To develop an efficient and flexible strategy for Drosophila melanogaster genome engineering.
- To combine CRISPR technology with recombinase-mediated cassette exchange (RMCE) for enhanced gene targeting.
- To establish a broadly applicable method for modifying multiple genomic locations in Drosophila.
Main Methods:
- A two-step strategy involving CRISPR-mediated integration followed by RMCE was developed.
- CRISPR co-injection with sgRNAs and a donor plasmid into Lig4 mutant embryos.
- Homologous integration identified by eye fluorescence, followed by RMCE for sequence replacement.
Main Results:
- The CRISPR-RMCE strategy demonstrated high efficiency in engineering four distinct genomic locations, including chromosome 4.
- The method allows for flexible gene modification by replacing an eye marker with desired DNA sequences.
- Successful application across multiple genomic sites suggests broad utility.
Conclusions:
- This efficient two-step CRISPR-RMCE strategy provides flexible genome engineering in Drosophila melanogaster.
- The method enables rapid modification of favorite genes for diverse research applications.
- This approach is expected to be accessible to any fly laboratory within approximately 3 months.
Related Concept Videos
02:53Conservative Site-specific Recombination and Phase Variation
5.7K
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...
5.7K
01:28CRISPR/Cas9 Genome Editing
3.0K
The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
3.0K