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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Insights into wild-type dynamin 2 and the consequences of DNM2 mutations from transgenic zebrafish.
TL;DR: Using the power of the zebrafish, novel insights are established into dynamin 2 localization and dynamics, and the necessary groundwork for future studies examining dynamin 1 pathomechanisms and therapy development is provided.
Journal ArticleDOI
MicroRNA-mediated control of developmental lymphangiogenesis
Hyun Min Jung,Ciara T Hu,Alexandra M Fister,Andrew M. Davis,Daniel Castranova,Van N. Pham,Lisa M Price,Brant M. Weinstein +7 more
TL;DR: High throughput small RNA sequencing is used to identify miR-204, a highly conserved microRNA dramatically enriched in lymphatic vs. blood endothelial cells in human and zebrafish, and shows that NFATC1 suppression leads to lymphatic hyperplasia, suggesting a critical positive role for this microRNA during developmental lymphangiogenesis.
Journal ArticleDOI
Conditional and biased regeneration of cone photoreceptor types in the zebrafish retina.
Florence D. D'Orazi,Florence D. D'Orazi,Sachihiro C. Suzuki,Sachihiro C. Suzuki,Nicole Darling,Nicole Darling,Rachel O.L. Wong,Takeshi Yoshimatsu,Takeshi Yoshimatsu +8 more
TL;DR: The regenerative capacity and stereotypic retinal organization of zebrafish was capitalized on to determine the specificity with which retinal Müller glial cells replace lost neuronal cell types, adding to the growing notion that cell replacement during regeneration does not perfectly mimic programs of cell generation during development.
Journal ArticleDOI
A high-conductance chemo-optogenetic system based on the vertebrate channel Trpa1b.
Pui-ying Lam,Suresh K. Mendu,Robert W. Mills,Baohui Zheng,Hugo Padilla,David J. Milan,Bimal N. Desai,Randall T. Peterson +7 more
TL;DR: The results demonstrate that zTrpa1b/ligand pairing offers high light sensitivity, millisecond-scale response latency in vivo, as well as adjustable channel off latency, and represents a novel tool for flexible, high-conductance optogenetics.
Journal ArticleDOI
Bsx controls pineal complex development.
TL;DR: The highly conserved homeodomain transcription factor Bsx controls zebrafish pineal gland development, laterality of the epithalamus and melatonin production.
References
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GAL4-VP16 is an unusually potent transcriptional activator
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Journal ArticleDOI
DNA Cloning Using In Vitro Site-Specific Recombination
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