Journal ArticleDOI
The Tol2kit: a multisite gateway-based construction kit for Tol2 transposon transgenesis constructs.
Kristen M. Kwan,Esther Fujimoto,Clemens Grabher,Benjamin D. Mangum,Melissa Hardy,Douglas Simon Campbell,John M. Parant,H. Joseph Yost,John P. Kanki,Chi Bin Chien +9 more
TLDR
The Tol2kit greatly facilitates zebrafish transgenesis, simplifies the sharing of clones, and enables large‐scale projects testing the functions of libraries of regulatory or coding sequences.Abstract:
Transgenesis is an important tool for assessing gene function. In zebrafish, transgenesis has suffered from three problems: the labor of building complex expression constructs using conventional subcloning; low transgenesis efficiency, leading to mosaicism in transient transgenics and infrequent germline incorporation; and difficulty in identifying germline integrations unless using a fluorescent marker transgene. The Tol2kit system uses site-specific recombination-based cloning (multisite Gateway technology) to allow quick, modular assembly of [promoter]-[coding sequence]-[3' tag] constructs in a Tol2 transposon backbone. It includes a destination vector with a cmlc2:EGFP (enhanced green fluorescent protein) transgenesis marker and a variety of widely useful entry clones, including hsp70 and beta-actin promoters; cytoplasmic, nuclear, and membrane-localized fluorescent proteins; and internal ribosome entry sequence-driven EGFP cassettes for bicistronic expression. The Tol2kit greatly facilitates zebrafish transgenesis, simplifies the sharing of clones, and enables large-scale projects testing the functions of libraries of regulatory or coding sequences.read more
Citations
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The histone methyltransferase SETDB1 is recurrently amplified in melanoma and accelerates its onset
Craig J. Ceol,Yariv Houvras,Yariv Houvras,Yariv Houvras,Judit Jané-Valbuena,Judit Jané-Valbuena,Steve Bilodeau,David A. Orlando,Valentine Battisti,Lauriane Fritsch,William M. Lin,William M. Lin,Travis J. Hollmann,Fabrizio Ferrè,Caitlin Bourque,Christopher J. Burke,Laura Turner,Audrey Uong,Laura A. Johnson,Laura A. Johnson,Rameen Beroukhim,Rameen Beroukhim,Craig H. Mermel,Craig H. Mermel,Massimo Loda,Slimane Ait-Si-Ali,Levi A. Garraway,Levi A. Garraway,Richard A. Young,Leonard I. Zon +29 more
TL;DR: SETDB1, an enzyme that methylates histone H3 on lysine 9 (H3K9), was found to accelerate melanoma formation significantly in zebrafish and is established as an oncogene in melanoma and underscores the role of chromatin factors in regulating tumorigenesis.
Journal ArticleDOI
Simultaneous single-cell profiling of lineages and cell types in the vertebrate brain.
Bushra Raj,Daniel E. Wagner,Aaron McKenna,Shristi Pandey,Allon M. Klein,Jay Shendure,Jay Shendure,James A. Gagnon,James A. Gagnon,Alexander F. Schier +9 more
TL;DR: ScGESTALT as discussed by the authors combines the lineage recording capabilities of GESTALT with cell-type identification by single-cell RNA sequencing, which relies on an inducible system that enables barcodes to be edited at multiple time points, capturing lineage information from later stages of development.
Journal ArticleDOI
MicroRNA-mediated integration of haemodynamics and Vegf signalling during angiogenesis
TL;DR: This work describes a novel genetic mechanism in which a microRNA facilitates integration of a physiological stimulus with growth factor signalling in endothelial cells to guide angiogenesis.
Journal ArticleDOI
Consensus guidelines for the use and interpretation of angiogenesis assays
Patrycja Nowak-Sliwinska,Kari Alitalo,Elizabeth Allen,Andrey Anisimov,Alfred C. Aplin,Robert Auerbach,Hellmut G. Augustin,Hellmut G. Augustin,David O. Bates,Judy R. van Beijnum,R. Hugh F. Bender,Gabriele Bergers,Gabriele Bergers,Andreas Bikfalvi,Joyce Bischoff,Barbara C. Böck,Barbara C. Böck,Peter C. Brooks,Federico Bussolino,Bertan Cakir,Peter Carmeliet,Daniel Castranova,Anca Maria Cimpean,Ondine Cleaver,George Coukos,George E. Davis,Michele De Palma,Anna Dimberg,Ruud P.M. Dings,Valentin Djonov,Andrew C. Dudley,Neil Dufton,Sarah-Maria Fendt,Napoleone Ferrara,Marcus Fruttiger,Dai Fukumura,Bart Ghesquière,Bart Ghesquière,Yan Gong,Robert J. Griffin,Adrian L. Harris,Christopher C.W. Hughes,Nan W. Hultgren,M. Luisa Iruela-Arispe,Melita Irving,Rakesh K. Jain,Raghu Kalluri,Joanna Kalucka,Robert S. Kerbel,Jan Kitajewski,Ingeborg Klaassen,Hynda K. Kleinmann,Pieter Koolwijk,Elisabeth Kuczynski,Brenda R. Kwak,Koen Marien,Juan M. Melero-Martin,Lance L. Munn,Roberto F. Nicosia,Agnès Noël,Jussi Nurro,Anna-Karin Olsson,Tatiana V. Petrova,Kristian Pietras,Roberto Pili,Jeffrey W. Pollard,Mark J. Post,Paul H.A. Quax,Gabriel A. Rabinovich,Marius Raica,Anna M. Randi,Domenico Ribatti,Curzio Rüegg,Reinier O. Schlingemann,Reinier O. Schlingemann,Stefan Schulte-Merker,Lois E.H. Smith,Jonathan W. Song,Steven A. Stacker,Jimmy Stalin,Amber N. Stratman,Maureen Van de Velde,Victor W.M. van Hinsbergh,Peter B. Vermeulen,Johannes Waltenberger,Brant M. Weinstein,Hong Xin,Bahar Yetkin-Arik,Seppo Ylä-Herttuala,Mervin C. Yoder,Arjan W. Griffioen +90 more
TL;DR: In vivo, ex vivo, and in vitro bioassays that are available for the evaluation of angiogenesis are described and critical aspects that are relevant for their execution and proper interpretation are highlighted.
Journal ArticleDOI
Ubiquitous transgene expression and Cre-based recombination driven by the ubiquitin promoter in zebrafish
Christian Mosimann,Charles K. Kaufman,Pulin Li,Pulin Li,Emily K. Pugach,Emily K. Pugach,Owen J. Tamplin,Owen J. Tamplin,Leonard I. Zon +8 more
TL;DR: The isolation and characterization of the zebrafish ubiquitin (ubi) promoter is reported, which drives constitutive transgene expression during all developmental stages and analyzed adult organs.
References
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Journal ArticleDOI
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Journal ArticleDOI
DNA Cloning Using In Vitro Site-Specific Recombination
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