GSBS Student Publications

Title

An enhanced gene targeting toolkit for Drosophila: Golic+

Student Author(s)

Hui-Min Chen

GSBS Program

Neuroscience

Publication Date

2015-3

UMMS Affiliation

Department of Neurobiology; Tzumin Lee Lab; Graduate School of Biomedical Sciences, Neuroscience Program

Document Type

Article

Disciplines

Genetics and Genomics | Neuroscience and Neurobiology

Abstract

Ends-out gene targeting allows seamless replacement of endogenous genes with engineered DNA fragments by homologous recombination, thus creating designer "genes" in the endogenous locus. Conventional gene targeting in Drosophila involves targeting with the preintegrated donor DNA in the larval primordial germ cells. Here we report G: ene targeting during O: ogenesis with L: ethality I: nhibitor and C: RISPR/Cas (Golic+), which improves on all major steps in such transgene-based gene targeting systems. First, donor DNA is integrated into precharacterized attP sites for efficient flip-out. Second, FLP, I-SceI, and Cas9 are specifically expressed in cystoblasts, which arise continuously from female germline stem cells, thereby providing a continual source of independent targeting events in each offspring. Third, a repressor-based lethality selection is implemented to facilitate screening for correct targeting events. Altogether, Golic+ realizes high-efficiency ends-out gene targeting in ovarian cystoblasts, which can be readily scaled up to achieve high-throughput genome editing.

DOI of Published Version

10.1534/genetics.114.173716

Source

Genetics. 2015 Mar;199(3):683-94. doi: 10.1534/genetics.114.173716. Epub 2015 Jan 2. Link to article on publisher's site

Journal/Book/Conference Title

Genetics

Related Resources

Link to Article in PubMed

PubMed ID

25555988

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