Dataset Information

An Interscholastic Network To Generate LexA Enhancer Trap Lines in Drosophila.

ABSTRACT: Binary expression systems like the LexA-LexAop system provide a powerful experimental tool kit to study gene and tissue function in developmental biology, neurobiology, and physiology. However, the number of well-defined LexA enhancer trap insertions remains limited. In this study, we present the molecular characterization and initial tissue expression analysis of nearly 100 novel StanEx LexA enhancer traps, derived from the StanEx1 index line. This includes 76 insertions into novel, distinct gene loci not previously associated with enhancer traps or targeted LexA constructs. Additionally, our studies revealed evidence for selective transposase-dependent replacement of a previously-undetected KP element on chromosome III within the StanEx1 genetic background during hybrid dysgenesis, suggesting a molecular basis for the over-representation of LexA insertions at the NK7.1 locus in our screen. Production and characterization of novel fly lines were performed by students and teachers in experiment-based genetics classes within a geographically diverse network of public and independent high schools. Thus, unique partnerships between secondary schools and university-based programs have produced and characterized novel genetic and molecular resources in Drosophila for open-source distribution, and provide paradigms for development of science education through experience-based pedagogy.

SUBMITTER: Kockel L

PROVIDER: S-EPMC6643891 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

An Interscholastic Network To Generate LexA Enhancer Trap Lines in Drosophila.

Kockel Lutz L Griffin Catherine C Ahmed Yaseen Y Fidelak Lauren L Rajan Arjun A Gould Ethan P EP Haigney Myles M Ralston Benjamin B Tercek Rex J RJ Galligani Lara L Rao Sagar S Huq Lutfi L Bhargava Hersh K HK Dooner Ailis C AC Lemmerman Emily G EG Malusa Ruby F RF Nguyen Tran H TH Chung Julie S JS Gregory Sara M SM Kuwana Kiyomasa M KM Regenold Jonathan T JT Wei Alexander A Ashton Jake J Dickinson Patrick P Martel Kate K Cai Connie C Chen Carissa C Price Stephen S Qiao Jeffrey J Shepley David D Zhang Joanna J Chalasani Meghana M Nguyen Khanh K Aalto August A Kim ByungJun B Tazawa-Goodchild Erik E Sherwood Amanda A Rahman Ahmad A Wu Sum Ying Celeste SYC Lotzkar Joel J Michaels Serena S Aristotle Hillary H Clark Antigone A Gasper Grace G Xiang Evan E Schlör Frieda Luna FL Lu Melissa M Haering Kate K Friberg Julia J Kuwana Alyssa A Lee Jonathan J Liu Alan A Norton Emma E Hamad Leena L Lee Clara C Okeremi Dara D diTullio Harry H Dumoulin Kat K Chi Sun Yu Gordon SYG Derossi Grayson S GS Horowitch Rose E RE Issa Elias C EC Le Dan T DT Morales Bryce C BC Noori Ayush A Shao Justin J Cho Sophia S Hoang Mai N MN Johnson Ian M IM Lee Katherine C KC Lee Maria M Madamidola Elizabeth A EA Schmitt Katrina E KE Byan Gabriel G Park Taeyoung T Chen Jonathan J Monovoukas Alexi A Kang Madison J MJ McGowan Tanner T Walewski Joseph J JJ Simon Brennan B Zu Sophia J SJ Miller Gregory P GP Fitzpatrick Kate B KB Lantz Nicole N Fox Elizabeth E Collette Jeanette J Kurtz Richard R Duncan Chris C Palmer Ryan R Rotondo Cheryl C Janicki Eric E Chisholm Townley T Rankin Anne A Park Sangbin S Kim Seung K SK

G3 (Bethesda, Md.) 20190709 7

Binary expression systems like the LexA-LexAop system provide a powerful experimental tool kit to study gene and tissue function in developmental biology, neurobiology, and physiology. However, the number of well-defined LexA enhancer trap insertions remains limited. In this study, we present the molecular characterization and initial tissue expression analysis of nearly 100 novel StanEx LexA enhancer traps, derived from the StanEx1 index line. This includes 76 insertions into ...[more]

PMID: 31040111

Similar Datasets

Project description:BACKGROUND: We have developed genetic methods in zebrafish by using the Tol2 transposable element; namely, transgenesis, gene trapping, enhancer trapping and the Gal4FF-UAS system. Gene trap constructs contain a splice acceptor and the GFP or Gal4FF (a modified version of the yeast Gal4 transcription activator) gene, and enhancer trap constructs contain the zebrafish hsp70l promoter and the GFP or Gal4FF gene. By performing genetic screens using these constructs, we have generated transgenic zebrafish that express GFP and Gal4FF in specific cells, tissues and organs. Gal4FF expression is visualized by creating double transgenic fish carrying a Gal4FF transgene and the GFP reporter gene placed downstream of the Gal4-recognition sequence (UAS). Further, the Gal4FF-expressing cells can be manipulated by mating with UAS effector fish. For instance, when fish expressing Gal4FF in specific neurons are crossed with the UAS:TeTxLC fish carrying the tetanus neurotoxin gene downstream of UAS, the neuronal activities are inhibited in the double transgenic fish. Thus, these transgenic fish are useful to study developmental biology and neurobiology. DESCRIPTION: To increase the usefulness of the transgenic fish resource, we developed a web-based database named zTrap http://kawakami.lab.nig.ac.jp/ztrap/. The zTrap database contains images of GFP and Gal4FF expression patterns, and genomic DNA sequences surrounding the integration sites of the gene trap and enhancer trap constructs. The integration sites are mapped onto the Ensembl zebrafish genome by in-house Blat analysis and can be viewed on the zTrap and Ensembl genome browsers. Furthermore, zTrap is equipped with the functionality to search these data for expression patterns and genomic loci of interest. zTrap contains the information about transgenic fish including UAS reporter and effector fish. CONCLUSION: zTrap is a useful resource to find gene trap and enhancer trap fish lines that express GFP and Gal4FF in desired patterns, and to find insertions of the gene trap and enhancer trap constructs that are located within or near genes of interest. These transgenic fish can be utilized to observe specific cell types during embryogenesis, to manipulate their functions, and to discover novel genes and cis-regulatory elements. Therefore, zTrap should facilitate studies on genomics, developmental biology and neurobiology utilizing the transgenic zebrafish resource.

Dataset Information

An Interscholastic Network To Generate LexA Enhancer Trap Lines in Drosophila.

Publications

An Interscholastic Network To Generate LexA Enhancer Trap Lines in <i>Drosophila</i>.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets