split-intein Gal4 provides intersectional genetic labeling that is fully repressible by Gal80

Ewen-Campen, Ben; Luan, Haojiang; Xu, Jun; Singh, Rohit; Joshi, Nitin; Thakkar, Tanuj; Berger, Bonnie; White, Benjamin H.; Perrimon, Norbert

doi:10.1101/2023.03.24.534001

Cited by 1 publication

(1 citation statement)

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“…Moreover, temporal regulation via AGES allows combining with other UAS transgenes for experiments like complementation analysis. Although the current version of AGES is incompatible with the many Gal80-insensitive split-Gal4 lines ( 25 ), it can be used with new Gal80-sensitive split-Gal4 lines ( 70 ). In summary, both temporal methods are effective, and we hope this exploration of their differences will help researchers choose the one best suited to their needs.…”

Section: Discussionmentioning

confidence: 99%

Dissecting neuron-specific functions of circadian genes using modified cell-specific CRISPR approaches

Richhariya

Shin

et al. 2023

Proc. Natl. Acad. Sci. U.S.A.

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Circadian behavioral rhythms in Drosophila melanogaster are regulated by about 75 pairs of brain neurons. They all express the core clock genes but have distinct functions and gene expression profiles. To understand the importance of these distinct molecular programs, neuron-specific gene manipulations are essential. Although RNAi based methods are standard to manipulate gene expression in a cell-specific manner, they are often ineffective, especially in assays involving smaller numbers of neurons or weaker Gal4 drivers. We and others recently exploited a neuron-specific CRISPR-based method to mutagenize genes within circadian neurons. Here, we further explore this approach to mutagenize three well-studied clock genes: the transcription factor gene vrille, the photoreceptor gene Cryptochrome ( cry ), and the neuropeptide gene Pdf (pigment dispersing factor). The CRISPR-based strategy not only reproduced their known phenotypes but also assigned cry function for different light-mediated phenotypes to discrete, different subsets of clock neurons. We further tested two recently published methods for temporal regulation in adult neurons, inducible Cas9 and the auxin-inducible gene expression system. The results were not identical, but both approaches successfully showed that the adult-specific knockout of the neuropeptide Pdf reproduces the canonical loss-of-function mutant phenotypes. In summary, a CRISPR-based strategy is a highly effective, reliable, and general method to temporally manipulate gene function in specific adult neurons.

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Section: Discussionmentioning

confidence: 99%