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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PatentDOI
BICISTRONIC GENE TRANSFER TOOLS FOR DELIVERY OF miRNAS AND PROTEIN CODING SEQUENCES
TL;DR: Results suggest that this cassette allows expression of miRNAs and proteins simultaneously, which provides the opportunity for joint delivery of specific translational repressors (miRNA) and possibly transcriptional activators (transcription factors) for future gene therapy use.
Journal ArticleDOI
Zebrafish: an underutilized tool for discovery in host-microbe interactions.
TL;DR: In this paper , the authors argue that the zebrafish model provides fertile ground for a new understanding of host-microbe interactions in a transparent host, and they use the model to study host-pathogen interactions.
Journal ArticleDOI
Dietary cholesterol and apolipoprotein A-I are trafficked in endosomes and lysosomes in the live zebrafish intestine.
Jessica P. Otis,Meng-Chieh Shen,Blake A. Caldwell,Oscar E. Reyes Gaido,Oscar E. Reyes Gaido,Steven A. Farber,Steven A. Farber +6 more
TL;DR: A novel assay to visualize live dietary cholesterol trafficking in the zebrafish intestine by feeding TopFluor-cholesterol, a fluorescent cholesterol analog, in a lipid-rich, chicken egg yolk feed is developed and elucidates a key role for endosomal-lysosomal trafficking of intestinal cholesterol and APOA-I.
Posted ContentDOI
Osmolarity-independent electrical cues guide rapid response to injury in zebrafish epidermis
TL;DR: It is found that, in addition to sensing osmolarity, basal skin cells in zebrafish larvae are also sensitive to changes in the particular ionic composition of their surroundings after wounding, specifically the concentration of sodium chloride in the immediate vicinity of the wound.
Journal ArticleDOI
Tfap2b specifies an embryonic melanocyte stem cell that retains adult multifate potential
TL;DR: In this article , Tfap2b and a select set of target genes specify melanocyte stem cell (McSC) population at the dorsal root ganglia in zebrafish.
References
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Journal ArticleDOI
DNA Cloning Using In Vitro Site-Specific Recombination
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