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
More filters
Posted ContentDOI
Fate plasticity and reprogramming in genetically distinct populations of Danio leucophores
Victor M Lewis,Lauren M. Saunders,Tracy A. Larson,Emily J. Bain,Samantha L. Sturiale,Dvir Gur,Sarwat Chowdhury,Jessica D. Flynn,Michael C. Allen,Dimitri D. Deheyn,Jennifer C. Lee,Julian A. Simon,Jennifer Lippincott-Schwartz,David W. Raible,David M. Parichy +14 more
TL;DR: Two distinct white pigment cell populations, leucophores, that arise from independent progenitors in zebrafish and its relatives are identified and contribute to the understanding of form and function in the spectacular pigment patterns of teleost fishes.
Journal ArticleDOI
Cell Adhesion in Zebrafish Embryos Is Modulated by March8
TL;DR: It is suggested that March8 functions in the embryo to modulate the strength of cell adhesion by regulating the localization of E-cadherin.
Journal ArticleDOI
uhrf1 and dnmt1 Loss Induces an Immune Response in Zebrafish Livers Due to Viral Mimicry by Transposable Elements.
Elena Magnani,Filippo Macchi,Bhavani P. Madakashira,Chi Zhang,Fatima Alaydaroos,Kirsten C. Sadler +5 more
TL;DR: In this paper, the authors provide mechanistic insight into the observation of an activated interferon response in the liver of zebrafish larvae with deletion in critical components of the DNA methylation machinery, uhrf1 and dnmt1.
Posted ContentDOI
Foxc1 establishes enhancer accessibility for craniofacial cartilage differentiation
TL;DR: It is proposed that Foxc1-dependent chromatin accessibility helps directs the versatile Sox9 protein to a chondrogenic program in the face by establishing accessibility in a subset of facial cartilage enhancers.
Journal ArticleDOI
Drug Screening to Treat Early-Onset Eye Diseases: Can Zebrafish Expedite the Discovery?
TL;DR: Visual-motor response, the activity response of larvae to a change in light stimuli, is proposed to serve as a simple and efficient tool for screening drugs that may improve vision in various zebrafish visual mutants.
References
More filters
Journal ArticleDOI
Stages of embryonic development of the zebrafish.
TL;DR: A series of stages for development of the embryo of the zebrafish, Danio (Brachydanio) rerio is described, providing for flexibility and continued evolution of the staging series as the authors learn more about development in this species.
Journal ArticleDOI
Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein.
Nathan C. Shaner,Robert E. Campbell,Robert E. Campbell,Paul Steinbach,Ben N G Giepmans,Amy E. Palmer,Roger Y. Tsien +6 more
TL;DR: The latest red version matures more completely, is more tolerant of N-terminal fusions and is over tenfold more photostable than mRFP1, and three monomers with distinguishable hues from yellow-orange to red-orange have higher quantum efficiencies.
Journal ArticleDOI
An Hsp70-like protein in the ER: identity with the 78 kd glucose-regulated protein and immunoglobulin heavy chain binding protein.
Sean Munro,Hugh R.B. Pelham +1 more
TL;DR: A cDNA clone is characterized that encodes a protein related to the 70 kd heat shock protein, but is expressed in normal rat liver, and it is identical with two previously described proteins: GRP78, whose synthesis is induced by glucose starvation, and BiP, which is found bound to immunoglobulin heavy chains in pre-B cells.
Journal ArticleDOI
GAL4-VP16 is an unusually potent transcriptional activator
TL;DR: It is shown that the hybrid protein (GAL4-VP16) activates transcription unusually efficiently in mammalian cells when bound close to, or at large distances from the gene, and suggested that the activating region of VP16 may be near-maximally potent.
Journal ArticleDOI
DNA Cloning Using In Vitro Site-Specific Recombination
TL;DR: A method called recombinational cloning is described that uses in vitro site-specific recombination to accomplish the directional cloning of PCR products and the subsequent automatic subcloning of the DNA segment into new vector backbones at high efficiency.