Targeting Endogenous Loci That Function in Drosophila Germline Stem Cells

Marianne Mercer; Varsha Bhargava; Courtney D Goldstein; Michael Buszczak

doi:10.1007/978-1-0716-3259-8_5

Targeting Endogenous Loci That Function in Drosophila Germline Stem Cells

Methods Mol Biol. 2023:2677:99-112. doi: 10.1007/978-1-0716-3259-8_5.

Authors

Marianne Mercer¹, Varsha Bhargava¹, Courtney D Goldstein¹, Michael Buszczak²

Affiliations

¹ Department of Molecular Biology, The University of Texas Southwestern Medical Center, Dallas, TX, USA.
² Department of Molecular Biology, The University of Texas Southwestern Medical Center, Dallas, TX, USA. Michael.buszczak@utsouthwestern.edu.

PMID: 37464237
DOI: 10.1007/978-1-0716-3259-8_5

Abstract

CRISPR-Cas9 genome editing technology can be used to manipulate the genome of Drosophila melanogaster. The ability to delete genes, make specific mutations, add tags, or make other genetic manipulations is useful for studying germline stem cell biology. In this chapter, we will describe a method to use CRISPR-Cas9 genome editing technology to make knock-out and knock-in flies. We will cover everything from guideRNA (gRNA) and donor plasmid design and cloning to screening for positive edits.

Keywords: CRISPR-Cas9; Drosophila ovary; Genome editing; Germline stem cells.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
CRISPR-Cas Systems
Drosophila melanogaster* / genetics
Drosophila* / genetics
Germ Cells
Stem Cells

Abstract

Publication types

MeSH terms

Grants and funding