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
Associative conditioning remaps odor representations and modifies inhibition in a higher olfactory brain area
Thomas Frank,Nila R Mönig,Nila R Mönig,Chie Satou,Shin-ichi Higashijima,Rainer W. Friedrich,Rainer W. Friedrich +6 more
TL;DR: It is found that a subregion in the zebrafish homolog of olfactory cortex maps odor space onto a representation of valence, which shapes this odor-to-valence map through plasticity processes that modify inhibition.
Journal ArticleDOI
Nr2f1b control venous specification and angiogenic patterning during zebrafish vascular development.
TL;DR: It is shown that nr2f1b control venous specification and angiogenic patterning during zebrafish vascular development, which is mediated by Notch signalings.
Journal ArticleDOI
Photoreceptor Progenitors Depend Upon Coordination of gdf6a, thrβ, and tbx2b to Generate Precise Populations of Cone Photoreceptor Subtypes.
Michèle G. DuVal,W. Ted Allison +1 more
TL;DR: The intersecting effects of cone development factors gdf6a, tbx2b, and thrβ are mapped in a model organism with high cone diversity, zebrafish, and it is revealed that disrupting thrβ activity did not change gDF6a mutant cone phenotypes.
Posted ContentDOI
High-throughput cellular-resolution synaptic connectivity mapping in vivo with concurrent two-photon optogenetics and volumetric Ca2+ imaging
Christopher McRaven,Dimitrii Tanese,Lixia Zhang,Chao-Tsung Yang,Misha B. Ahrens,Valentina Emiliani,Minoru Koyama +6 more
TL;DR: A circuit motif for generating fast undulatory locomotion in zebrafish is revealed by combining two-photon optogenetics and volumetric Ca2+ imaging with whole-cell recording for high-throughput synaptic connectivity and properties measurements.
Journal ArticleDOI
Loss of Pde6 reduces cell body Ca2+ transients within photoreceptors
TL;DR: In vivo imaging strategies based on genetically encoded Ca2+ indicators are exploited and a dramatic, light-independent modulation of [Ca2+]i that occurs in normal cones is uncovered and provides insight into Ca2- dynamics in a common form of inherited blindness.
References
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Journal ArticleDOI
GAL4-VP16 is an unusually potent transcriptional activator
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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.