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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Journal ArticleDOI
Cavin4 interacts with Bin1 to promote T-tubule formation and stability in developing skeletal muscle.
Harriet P. Lo,Ye-Wheen Lim,Zherui Xiong,Nick Martel,Charles Ferguson,Nicholas Ariotti,Jean Giacomotto,James Rae,Matthias Floetenmeyer,Shayli Varasteh Moradi,Ya Gao,Vikas A. Tillu,Di Xia,Huang Wang,Samira Rahnama,Susan J. Nixon,Michele Bastiani,Ryan D. Day,Kelly A. Smith,Nathan J. Palpant,Wayne A. Johnston,Kirill Alexandrov,Brett M. Collins,Thomas E. Hall,Robert G. Parton +24 more
TL;DR: In this paper, a role for muscle-specific cavin proteins in skeletal muscle T-tubule development was identified by analyzing two vertebrate systems, mouse and zebrafish.
Journal Article
Genetic Analysis of the Touch Response in Zebrafish (Danio rerio).
TL;DR: The advantages of zebrafish are reviewed for studies of the biological mechanisms involved in touch sensitivity and it is demonstrated that a simple touch assay efficiently recovers mutations that affect sensory neurons.
Journal ArticleDOI
Adaptive cell invasion maintains lateral line organ homeostasis in response to environmental changes.
Julia Peloggia,Daniela Münch,Paloma Meneses-Giles,Andrés Romero-Carvajal,Andrés Romero-Carvajal,Mark E. Lush,Nathan D. Lawson,Melainia McClain,Y. Albert Pan,Y. Albert Pan,Tatjana Piotrowski +10 more
TL;DR: In this paper, the authors used lineage labeling and scRNA-seq to identify Neuromast-associated (Nm) ionocytes, which are likely regulators of sensory hair cells.
Posted ContentDOI
Identification of 3’ UTR motifs required for mRNA localization to myelin sheaths in vivo
TL;DR: It is shown that one motif is highly enriched in the myelin transcriptome, suggesting that this sequence is a global regulator of mRNA localization during developmental myelination.
Journal ArticleDOI
Development of a method to monitor gene expression in single bacterial cells during the interaction with plants and use to study the expression of the type III secretion system in single cells of Dickeya dadantii in potato
Zhouqi Cui,Xiaochen Yuan,Ching-Hong Yang,Regan B. Huntley,Weimin Sun,Jie Wang,George W. Sundin,Quan Zeng +7 more
TL;DR: A method that enabled in situ observation and quantification of gene expression in single bacterial cells in planta and observed that the expression of the T3SS genes hrpA and hrpN is restricted to a small proportion of D. dadantii cells during the infection of potato is developed.
References
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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.