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HAMA Vibes Optical Mouse Driver
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Received Feb 27; Accepted May 2. This article has been cited by other articles in PMC. Zebrafish, Signal, Pathway, Transgenic, Biosensor, Reporter Introduction Zebrafish Danio rerio is a small teleost that can be maintained in small aquaria.
The main advantage of working with zebrafish is that it is an animal with all characteristics of vertebrates and an organism with external HAMA Vibes Optical Mouse. These features, coupled with the fact that its embryo is semitransparent during early life stages, open the possibility to follow early development and organogenesis directly in vivo, without the need for invasive procedures.
Moreover, zebrafish are particularly fertile and a healthy adult female can lay hundreds of eggs in a month. For these reasons, zebrafish is the first vertebrate model in which large-scale random mutagenesis could be undertaken Driever et al.
The zebrafish genome has recently been sequenced and assembled, but its annotation is still ongoing. Moreover, until few years ago, genetic manipulation technology lacked a direct tool for targeting the zebrafish genome, but recently techniques have evolved through the use of TILLING McCallum et al.
Being a vertebrate, zebrafish HAMA Vibes Optical Mouse immediately gained acceptance as a model organism in which human diseases can be phenocopied. In fact, many human diseases can be modeled in zebrafish and several laboratories all over the world are now generating fish with diseases that resemble, either at the molecular, cellular, etiological or histopathological level, human diseases.
Several zebrafish models of cancer, metabolic, developmental, aging, degenerative, sensory organ and infectious diseases have been generated reviewed in Lieschke and Currie ; Mione and Trede ; Kanther and Rawls ; Liu and HAMA Vibes Optical Mouse ; Goldsmith and Jobin This scenario makes zebrafish an ideal candidate for screening disease modifiers, genes, drugs or behavioral and environmental mechanisms able to modify specific phenotypes.
The knowledge of molecular and biochemical processes regulating cell proliferation, differentiation, survival, and death is the basis for understanding animal development, tissue homeostasis and tumor growth. Many embryonic and postnatal signaling pathways, which are often disrupted in cancer and diseases, appear to play a central role.
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Particularly important in regulating cell proliferation, stem cell maintenance and differentiation are bone morphogenetic proteins BMPsnotch, wingless-related integration site Wntsonic Hedgehog Shhfibroblast growth factor HAMA Vibes Optical Mousesignal transducer and activator of transcription STATactivator protein 1 AP1 and cAMP-response element-binding protein CREB signaling reviewed in Perrimon et al. In fact, research at the tissue and cellular level is deeply oriented toward dissection of these pathways, as well as understanding their cross-talk and epistatic relations Davidson ; Corson and Siggia Typically, the analysis of genetic cascades culminates in dissecting cis-elements in the promoters of relevant target genes and the subsequent isolation of the TFs binding to these cis-elements.
This approach leads to the isolation of relevant TFs in genetic cascades and the exact knowledge of the activator sequences in the promoters and enhancers of target genes reviewed by Khalil et al. The direct and final proof that a specific TF and HAMA Vibes Optical Mouse the upstream genetic cascade is involved in gene activation through a specific nucleotide sequence cis-element comes from the strong and tunable activation of synthetic constructs bearing multimerized copies of the cis-element bound to a minimal promoter of a reporter gene.
In this way, a huge amount of data have been produced by transfection of a specific cell line together with the reporter and components of the signal transduction cascade.
However, this approach might lack in vivo relevance, as co-transfection of a reporter plasmid together with its transduction and TFs leads, by HAMA Vibes Optical Mouse, to reporter signal. For this reason, a number of transgenic animal reporter lines have been developed, from mice to Drosophila and, recently, zebrafish. In this review, we will focus on the advantage of using zebrafish in this approach, to convince the reader that embryo and larval transparency, genetic and chemical manipulation, screening properties and transgenic manipulability, make zebrafish an HAMA Vibes Optical Mouse tool for analytical screenings and analysis of pathway reporters.
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Development of signaling pathway reporters SPRs The generation of reliable transgenic fish expressing in vivo reporter proteins under the control of signaling pathway responsive elements depends on four major critical issues: Since the first successful attempt to introduce foreign DNA into the zebrafish germline Stuart et al. The initial method of transgenesis in zebrafish entailed the microinjection of linearized HAMA Vibes Optical Mouse into fertilized eggs.
However, this approach suffered from many drawbacks, including low rate of germline transmission and mosaic expression of the introduced transgene. Despite its limited yield and other minor issues, the first Wnt reporter line was established in the laboratory of Randall Moon in using this method Dorsky et al. This method relies on the generation of a targeting sequence flanked by two recognition sites for the I-SceI enzyme, an intron-encoded homing endonuclease isolated from the yeast Saccharomyces cerevisiae.
Co-injection of the vector with the I-SceI meganuclease significantly increases the rate of germline integration, thus improving the frequency of positive founder fish in F0. The development of the Tol2 technology has revolutionized transgenesis methods HAMA Vibes Optical Mouse aquatic vertebrates, such as zebrafish Kawakami et al. This method relies on the microinjection into fertilized eggs of a transposon-donor plasmid named Tol2, and a synthetic HAMA Vibes Optical Mouse encoding the transposase, to catalyze DNA excision and recombination within the targeted genome.
The Tol2 transgenesis method has been settled in most zebrafish laboratories worldwide and, in the past few years, many cell signaling reporter fish have been established through the application of this method Schwend et al.