Transgenic <i>Drosophila</i> lines for LexA-dependent gene and growth regulation

Chang, Kathleen; Tsao, Deborah D.; Bennett, Celine; Wang, Elaine; Floyd, Jax; Greenwald, Emily; Kim, Ella S; Griffin, Catherine; Morse, Elizabeth; Chisholm, Townley; Rankin, Anne E; Lopez, Luis Alberto Baena; Lantz, Nicole; Fox, Elizabeth; Kockel, Lutz; Kim, Seung K.; Park, Sangbin

doi:10.1101/2021.10.07.463520

Cited by 2 publications

(2 citation statements)

References 26 publications

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“…CRISPR/Cas9 technology now makes it possible to knock-in a transcriptional activator into any locus. Some groups have also recently described tools to swap QF or LexA into the GAL4 coding region of existing GAL4 enhancer trap lines (Lin and Potter 2016;Chang et al 2022;Karuparti et al 2023). The conversion can be performed through genetic crosses; however, the frequency of conversion can vary greatly among different Gal4 lines, and many such swaps do not fully reproduce the original GAL4 expression patterns (Chen et al 2019).…”

Section: Rapid and Efficient Generation Of Driver Lines By Crispr-cas9mentioning

confidence: 99%

Expanding the Drosophila toolkit for dual control of gene expression

Zirin,

Jusiak,

Lopes

et al. 2023

Preprint

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Having tools and resources for independent control of gene expression in two different tissues in the same animal is emerging as a major need, especially in the context of inter-organ communication studies. This type of study is made possible by technologies combining the GAL4/UAS and a second binary expression system such as LexA/LexAop or QF/QUAS. Here, we describe a resource of reagents that facilitate combined use of the GAL4/UAS and a second binary systems in various tissues. Focusing on genes with well-characterized GAL4 expression patterns, we generated a set of more than 40 LexA-GAD and QF2 insertions by CRISPR knock-in and verified their tissue-specificity in larvae. We also built constructs that encode QF2 and LexA-GAD transcription factors in a single vector, with each coding sequence flanked by FRT sites. Following successful integration into the fly genome, the vector generates a single polypeptide, which is separated into the individual drivers by the inclusion of T2A. If desired, one of the two coding regions can then be excised with Flp, resulting in flies that express only QF2 or LexA-GAD. We evaluated both QF2 and LexA-GAD systems for in vivo gene knockdown and are generating a compatible library of transgenic shRNA lines in our custom QUAS and LexAop vectors as a community resource. Together, these QF2/LexA-GAD and QUAS/LexAop vectors and fly lines will provide a new set of tools for researchers who need to activate or repress two different genes in an orthogonal manner in the same animal.

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Section: Rapid and Efficient Generation Of Driver Lines By Crispr-cas9mentioning

confidence: 99%

Expanding the Drosophila toolkit for dual control of gene expression

Zirin,

Jusiak,

Lopes

et al. 2023

Preprint

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“…In our prior studies, we produced and characterized novel fly enhancer trap lines through an interscholastic partnership of secondary school and university-based researchers in the U.S. (Kockel et al 2016(Kockel et al , 2019Chang et al 2022). This involved development of curricula permitting flexible scheduling of laboratory-based 'modules': in some schools, this occurred in year-long courses (Suppl.…”

Section: Somatic Expression Of Natural Transposonsmentioning

confidence: 99%

An international scholastic network to generate LexA enhancer-trap lines forDrosophila

Kim

Rajan

Chang

et al. 2022

Preprint

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Conditional gene regulation in Drosophila through binary expression systems like the LexA-LexAop system provides a superb tool for investigating gene and tissue function. To increase the availability of defined LexA enhancer trap insertions, we present molecular, genetic and tissue expression studies of 301 novel Stan-X LexA enhancer traps derived from a screen with the index SX4 line. This includes insertions into distinct loci on the X, II and III chromosomes that were not previously associated with enhancer traps or targeted LexA constructs. A subset of enhancer traps was expressed in CNS neurons known to produce and secrete insulin, an essential regulator of growth, development and metabolism. Fly lines described here were generated and characterized through studies by students and teachers in an international network of genetics classes at public and independent high schools and universities serving a diversity of students, including those underrepresented in science. Thus, a unique partnership between secondary schools and university-based programs has produced and characterized novel resources in Drosophila, establishing instructional paradigms devoted to unscripted experimental science.

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Transgenic Drosophila lines for LexA-dependent gene and growth regulation

Cited by 2 publications

References 26 publications

Expanding the Drosophila toolkit for dual control of gene expression

Expanding the Drosophila toolkit for dual control of gene expression

An international scholastic network to generate LexA enhancer-trap lines forDrosophila

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