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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ATX-LPA1 axis contributes to proliferation of chondrocytes by regulating fibronectin assembly leading to proper cartilage formation.
Tatsuji Nishioka,Naoaki Arima,Kuniyuki Kano,Kotaro Hama,Eriko Itai,Hiroshi Yukiura,Ryoji Kise,Asuka Inoue,Asuka Inoue,Seok-Hyung Kim,Lilianna Solnica-Krezel,Wouter H. Moolenaar,Jerold Chun,Junken Aoki,Junken Aoki +14 more
TL;DR: It is shown here that loss of ATX-LPA1 signaling leads to disorganization of chondrocytes, causing severe defects in cartilage formation, and the ATx- LPA1 axis is a key regulator of Cartilage formation.
Journal ArticleDOI
3-OST-7 Regulates BMP-Dependent Cardiac Contraction
Shiela C. Samson,Tania Ferrer,Chuanchau J. Jou,Frank B. Sachse,Sunita S. Shankaran,Robin M. Shaw,Neil C. Chi,Martin Tristani-Firouzi,H. Joseph Yost +8 more
TL;DR: During zebrafish cardiac development, 3-OST-7 constrains BMP signaling to the atrioventricular junction and precludes it from contractile myocardium, allowing tropomyosin-dependent sarcomere assembly and contraction.
Journal ArticleDOI
Anosmin1 Shuttles Fgf to Facilitate Its Diffusion, Increase Its Local Concentration, and Induce Sensory Organs
John Wang,Yandong Yin,Stephanie Lau,Jagadish Sankaran,Eli Rothenberg,Thorsten Wohland,Martin Meier-Schellersheim,Holger Knaut +7 more
TL;DR: The diffusion behavior of Fgf10 molecules during sensory organ formation in the zebrafish posterior lateral line primordium is analyzed and Anosmin1, which when mutated in humans causes Kallmann Syndrome, is identified as an ECM protein that binds to F gf10 and facilitates its diffusivity by increasing the pool of fast-moving FgF10 molecules.
Journal ArticleDOI
Long-Range Optogenetic Control of Axon Guidance Overcomes Developmental Boundaries and Defects
James M. Harris,Andy Yu-Der Wang,Jonathan Boulanger-Weill,Cristina Santoriello,Stephan Foianini,Jeff W. Lichtman,Leonard I. Zon,Leonard I. Zon,Paola Arlotta,Paola Arlotta +9 more
TL;DR: Zebrafish motor neurons expressing a photoactivatable Rac1 are used to co-opt endogenous growth cone guidance machinery to precisely and non-invasively direct axon growth using light, demonstrating that intrinsic growthcone guidance machinery can be co-opted to non- invasively build new connectivity, allowing investigation of neural network dynamics in intact living organisms.
Journal ArticleDOI
The Novel Small Molecule TRVA242 Stabilizes Neuromuscular Junction Defects in Multiple Animal Models of Amyotrophic Lateral Sclerosis
Poulomee Bose,Elsa Tremblay,Claudia Maios,Vijay Narasimhan,Gary A. B. Armstrong,Meijiang Liao,J. Alex Parker,Richard Robitaille,Xiao-Yan Wen,Christopher Barden,Pierre Drapeau +10 more
TL;DR: TRVA242 was identified as the most potent compound as it significantly improved efficiency in rescuing locomotor, motorneuron, and neuromuscular junction synaptic deficits in a C. elegans and genetic zebrafish models of ALS.
References
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Journal ArticleDOI
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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.
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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.