Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Genetic Screens02:46

Genetic Screens

5.0K
Genetic screens are tools used to identify genes and mutations responsible for phenotypes of interest. Genetic screens help identify individuals or a group of people at risk of developing  genetic diseases and help them with early intervention, targeted therapy, and reproductive options.
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which...
5.0K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

A multiplex hextuple luciferase assay for advancing cellular signaling analysis in plant cells.

Plant communications·2026
Same author

Synthetic Assembly DNA Cloning to Build Plasmids for Multiplexed Transgenic Selection, Counterselection or Any Other Genetic Strategies Using Drosophila melanogaster.

Current protocols·2023
Same author

Serial Recombineering Cloning to Build Selectable and Tagged Genomic P[acman] BAC Clones for Selection Transgenesis and Functional Gene Analysis using Drosophila melanogaster.

Current protocols·2023
Same author

The need for unbiased genetic screens to dissect aggression in <i>Drosophila melanogaster</i>.

Frontiers in behavioral neuroscience·2022
Same author

Synthetic Assembly DNA Cloning of Multiplex Hextuple Luciferase Reporter Plasmids.

Methods in molecular biology (Clifton, N.J.)·2022
Same author

Multiplex Hextuple Luciferase Assaying.

Methods in molecular biology (Clifton, N.J.)·2022

Related Experiment Video

Updated: Aug 11, 2025

Author Spotlight: Optimizing Embryo Microinjection for Transgenesis in Drosophila
05:32

Author Spotlight: Optimizing Embryo Microinjection for Transgenesis in Drosophila

Published on: June 7, 2024

2.2K

Multiplexed Transgenic Selection and Counterselection Strategies to Expedite Genetic Manipulation Workflows Using

Koen J T Venken1,2,3,4, Nick Matinyan1,3,5, Yezabel Gonzalez1

  • 1Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas.

Current Protocols
|February 9, 2023
PubMed
Summary

New drug-based markers in Drosophila melanogaster simplify genetic manipulation. These selectable and counterselectable markers streamline the process of creating transgenic animals for research, eliminating laborious screening steps.

Keywords:
Drosophila melanogastercounterselectiongenetic manipulationgeneticsmultiplexedselectiontransgenesis

More Related Videos

An Efficient Strategy for Generating Tissue-specific Binary Transcription Systems in Drosophila by Genome Editing
10:01

An Efficient Strategy for Generating Tissue-specific Binary Transcription Systems in Drosophila by Genome Editing

Published on: September 19, 2018

9.1K
One-step CRISPR-based Strategy for Endogenous Gene Tagging in Drosophila melanogaster
07:23

One-step CRISPR-based Strategy for Endogenous Gene Tagging in Drosophila melanogaster

Published on: January 26, 2024

843

Related Experiment Videos

Last Updated: Aug 11, 2025

Author Spotlight: Optimizing Embryo Microinjection for Transgenesis in Drosophila
05:32

Author Spotlight: Optimizing Embryo Microinjection for Transgenesis in Drosophila

Published on: June 7, 2024

2.2K
An Efficient Strategy for Generating Tissue-specific Binary Transcription Systems in Drosophila by Genome Editing
10:01

An Efficient Strategy for Generating Tissue-specific Binary Transcription Systems in Drosophila by Genome Editing

Published on: September 19, 2018

9.1K
One-step CRISPR-based Strategy for Endogenous Gene Tagging in Drosophila melanogaster
07:23

One-step CRISPR-based Strategy for Endogenous Gene Tagging in Drosophila melanogaster

Published on: January 26, 2024

843

Area of Science:

  • Genetics and Genomics
  • Molecular Biology
  • Developmental Biology

Background:

  • Traditional methods for identifying desired genotypes in Drosophila melanogaster involve laborious screening of progeny.
  • Visual or fluorescent markers require extensive analysis, hindering efficient genetic manipulation.
  • A need exists for streamlined selection strategies in Drosophila transgenesis.

Purpose of the Study:

  • To describe detailed procedures for utilizing drug-based selectable and counterselectable markers in Drosophila melanogaster.
  • To demonstrate the application of these markers for generating dual transgenic animals for binary overexpression systems.
  • To provide protocols for preparing drugged food, determining drug concentrations, and performing microinjection and crossing schemes.

Main Methods:

  • Development and application of four selectable markers (resistance to G418 sulfate, puromycin HCl, blasticidin S, hygromycin B) and two counterselectable markers (sensitivity to ganciclovir/acyclovir, 5-fluorocytosine).
  • Co-injection of plasmids encoding GAL4 transactivator and UAS responder for dual transgenesis.
  • Co-selection using specific drug combinations to isolate co-transgenic flies.
  • Detailed protocols for fly food preparation, drug concentration determination, plasmid preparation, microinjection, and crossing schemes.

Main Results:

  • Successful generation of dual transgenic Drosophila melanogaster for binary overexpression systems using co-injection and co-selection.
  • Demonstration of marker functionality through expression pattern analysis of dual transgenics.
  • Established protocols for implementing drug-based selection and counterselection strategies in Drosophila research.

Conclusions:

  • Drug-based selectable and counterselectable markers offer a significant advancement over traditional screening methods in Drosophila.
  • These markers facilitate efficient generation of dual transgenic animals, balancer chromosomes, and BAC transgenic lines.
  • The described protocols are adaptable for various marker combinations and genetic strategies, enhancing research flexibility.