Next-generation plasmids for transgenesis in zebrafish and beyond

Author:

Kemmler Cassie L.1ORCID,Moran Hannah R.1ORCID,Murray Brooke F.2ORCID,Scoresby Aaron2,Klem John R.3,Eckert Rachel L.3,Lepovsky Elizabeth3,Bertho Sylvain4,Nieuwenhuize Susan15,Burger Sibylle5,D'Agati Gianluca5ORCID,Betz Charles6ORCID,Puller Ann-Christin5,Felker Anastasia5ORCID,Ditrychova Karolina5ORCID,Bötschi Seraina5,Affolter Markus6ORCID,Rohner Nicolas4ORCID,Lovely C. Ben3ORCID,Kwan Kristen M.2ORCID,Burger Alexa1ORCID,Mosimann Christian1ORCID

Affiliation:

1. University of Colorado, School of Medicine, Anschutz Medical Campus 1 , Department of Pediatrics, Section of Developmental Biology, 12801 E 17th Avenue, Aurora, CO 80045 , USA

2. University of Utah 2 Department of Human Genetics , , Salt Lake City, UT 84112 , USA

3. University of Louisville School of Medicine 3 Department of Biochemistry and Molecular Genetics , , Louisville, KY 40202 , USA

4. Stowers Institute for Medical Research 4 , Kansas City, MO 64110 , USA

5. University of Zurich 5 Department of Molecular Life Sciences , , 8057 Zürich , Switzerland

6. University of Basel 6 Growth & Development, Biozentrum, Spitalstrasse 41 , , 4056 Basel , Switzerland

Abstract

ABSTRACT Transgenesis is an essential technique for any genetic model. Tol2-based transgenesis paired with Gateway-compatible vector collections has transformed zebrafish transgenesis with an accessible modular system. Here, we establish several next-generation transgenesis tools for zebrafish and other species to expand and enhance transgenic applications. To facilitate gene regulatory element testing, we generated Gateway middle entry vectors harboring the small mouse beta-globin minimal promoter coupled to several fluorophores, CreERT2 and Gal4. To extend the color spectrum for transgenic applications, we established middle entry vectors encoding the bright, blue-fluorescent protein mCerulean and mApple as an alternative red fluorophore. We present a series of p2A peptide-based 3′ vectors with different fluorophores and subcellular localizations to co-label cells expressing proteins of interest. Finally, we established Tol2 destination vectors carrying the zebrafish exorh promoter driving different fluorophores as a pineal gland-specific transgenesis marker that is active before hatching and through adulthood. exorh-based reporters and transgenesis markers also drive specific pineal gland expression in the eye-less cavefish (Astyanax). Together, our vectors provide versatile reagents for transgenesis applications in zebrafish, cavefish and other models.

Funder

National Institutes of Health

National Institute of Diabetes and Digestive and Kidney

University of Colorado School of Medicine, Anschutz Medical Campus

Children's Hospital Colorado

National Science Foundation

National Eye Institute

National Institute on Alcohol Abuse and Alcoholism

National Institute of General Medical Sciences

Swiss Bridge Foundation

Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung

Stowers Institute for Medical Research

Publisher

The Company of Biologists

Subject

Developmental Biology,Molecular Biology

同舟云学术

1.学者识别学者识别

2.学术分析学术分析

3.人才评估人才评估

"同舟云学术"是以全球学者为主线,采集、加工和组织学术论文而形成的新型学术文献查询和分析系统,可以对全球学者进行文献检索和人才价值评估。用户可以通过关注某些学科领域的顶尖人物而持续追踪该领域的学科进展和研究前沿。经过近期的数据扩容,当前同舟云学术共收录了国内外主流学术期刊6万余种,收集的期刊论文及会议论文总量共计约1.5亿篇,并以每天添加12000余篇中外论文的速度递增。我们也可以为用户提供个性化、定制化的学者数据。欢迎来电咨询!咨询电话:010-8811{复制后删除}0370

www.globalauthorid.com

TOP

Copyright © 2019-2024 北京同舟云网络信息技术有限公司
京公网安备11010802033243号  京ICP备18003416号-3