Determining Effective Drug Concentrations for Selection and Counterselection Genetics in

Overview

Journal STAR Protoc

Publisher Cell Press

Specialties Biology
Biomedical Engineering
Science

Date 2021 Sep 29

PMID 34585147

Citations 5

Authors

Nick Matinyan

Yezabel Gonzalez

Herman A Dierick

Koen J T Venken

Affiliations

Soon will be listed here.

Abstract

We recently integrated into fly genetics a set of four selection and two counterselection markers and their corresponding drugs that can be used individually or in combination. These markers eliminate the need to visually screen progeny. Before using these markers in new genetic backgrounds, effective selection/counterselection concentrations should be established for each marker/drug combination. This protocol describes how to set up, perform, and analyze a drug titration curve to determine the effective selection/counterselection drug concentrations for their corresponding markers. For complete details on the use and execution of this protocol, please refer to Matinyan et al., 2021.

Citing Articles

Redesigning the Drosophila histone gene cluster: an improved genetic platform for spatiotemporal manipulation of histone function.

Crain A, Nevil M, Leatham-Jensen M, Reeves K, Matera A, McKay D Genetics. 2024; 228(1).

PMID: 39039029 PMC: 11373521. DOI: 10.1093/genetics/iyae117.

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

Venken K, Matinyan N, Gonzalez Y, Dierick H Curr Protoc. 2023; 3(2):e675.

PMID: 36757632 PMC: 9923880. DOI: 10.1002/cpz1.675.

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

Venken K, Matinyan N, Gonzalez Y, Sarrion-Perdigones A, Dierick H Curr Protoc. 2023; 3(2):e653.

PMID: 36757602 PMC: 10281009. DOI: 10.1002/cpz1.653.

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

Venken K, Matinyan N, Gonzalez Y, Dierick H Curr Protoc. 2023; 3(2):e652.

PMID: 36757287 PMC: 9923875. DOI: 10.1002/cpz1.652.

Multiplexed drug-based selection and counterselection genetic manipulations in Drosophila.

Matinyan N, Karkhanis M, Gonzalez Y, Jain A, Saltzman A, Malovannaya A Cell Rep. 2021; 36(11):109700.

PMID: 34525356 PMC: 8480232. DOI: 10.1016/j.celrep.2021.109700.

References

Steller H, Pirrotta V . A transposable P vector that confers selectable G418 resistance to Drosophila larvae. EMBO J. 1985; 4(1):167-71. PMC: 554166. DOI: 10.1002/j.1460-2075.1985.tb02332.x. View

Venken K, Bellen H . Transgenesis upgrades for Drosophila melanogaster. Development. 2007; 134(20):3571-84. DOI: 10.1242/dev.005686. View

Kandul N, Liu J, Hsu A, Hay B, Akbari O . A drug-inducible sex-separation technique for insects. Nat Commun. 2020; 11(1):2106. PMC: 7193620. DOI: 10.1038/s41467-020-16020-2. View

Matinyan N, Karkhanis M, Gonzalez Y, Jain A, Saltzman A, Malovannaya A . Multiplexed drug-based selection and counterselection genetic manipulations in Drosophila. Cell Rep. 2021; 36(11):109700. PMC: 8480232. DOI: 10.1016/j.celrep.2021.109700. View