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Genesis: the Journal of Genetics and Development

Nicolas Unsain, Genevieve Dorval, JaeHyung Sheen, Philip A Barker
The Neurotrophin receptor associated death domain gene (Nradd/Nrh2/Plaidd) is a type I transmembrane protein with a unique and short N-terminal extracellular domain and a transmembrane and intracellular domain that bears high similarity to the p75 neurotrophin receptor (p75NTR/Ngfr). Initial studies suggested that NRADD regulates neurotrophin signaling but very little is known about its physiological roles. We have generated and characterized NRADD conditional and germ-line null mouse lines. These mice are viable and fertile and dońt show evident abnormalities...
October 24, 2016: Genesis: the Journal of Genetics and Development
Michael S Samuel, Nicola Rath, Siti F Masre, Sarah T Boyle, David A Greenhalgh, Marina Kochetkova, Sheila Bryson, David Stevenson, Michael F Olson
The serine/threonine kinases ROCK1 and ROCK2 are central mediators of actomyosin contractile force generation that act downstream of the RhoA small GTP-binding protein. As a result, they have key roles in regulating cell morphology and proliferation, and have been implicated in numerous pathological conditions and diseases including hypertension and cancer. Here we describe the generation of a gene-targeted mouse line that enables CRE-inducible expression of a conditionally-active fusion between the ROCK2 kinase domain and the hormone-binding domain of a mutated estrogen receptor (ROCK2:ER)...
October 24, 2016: Genesis: the Journal of Genetics and Development
Stefan Thiem, Moritz F Eissmann, Emma Stuart, Joachim Elzer, Anna Jonas, Michael Buchert, Matthias Ernst
Temporal and spatial regulation of genes mediated by tissue-specific promoters and conditional gene expression systems provide a powerful tool to study gene function in health, disease and during development. Although transgenic mice expressing the Cre recombinase in the gastric epithelium have been reported, there is a lack of models that allow inducible and reversible gene modification in the stomach. Here, we exploited the gastrointestinal epithelium-specific expression pattern of the three trefoil factor (Tff) genes and bacterial artificial chromosome transgenesis to generate a novel mouse strain that expresses the CreERT2 recombinase and the reverse tetracycline transactivator (rtTA)...
October 12, 2016: Genesis: the Journal of Genetics and Development
Huizhen Huang, Marissa S Kuzirian, Xiaoyun Cai, Lindsey M Snyder, Jonathan Cohen, Daniel H Kaplan, Sarah E Ross
The Neurokinin 1 Receptor (NK1R), which binds Substance P, is expressed in discrete populations of neurons throughout the nervous system, where it has numerous roles including the modulation of pain and affective behaviors. Here, we report the generation of a NK1R-CreER knockin allele, in which CreER(T2) replaces the coding sequence of the TACR1 gene (encoding NK1R) in order to gain genetic access to these cells. We find that the NK1R-CreER allele mediates recombination in many regions of the nervous system that are important in pain and anxiety including the amygdala, hypothalamus, frontal cortex, raphe nucleus, and dorsal horn of the spinal cord...
October 6, 2016: Genesis: the Journal of Genetics and Development
Lisa E Kelly, Reyna I Martinez-De Luna, Heithem M El-Hodiri
The Retinal homeobox (rax) gene is expressed in vertebrate retinal progenitor and stem cells and is essential for retinal development. In frogs, rax is expressed in the ciliary marginal zone (CMZ), a region containing retinal progenitor and stem cells at the anterior of the eye. Little is known regarding regulation of rax transcription and regulation of transcription of rax targets. We found that three ultra-conserved genomic elements (UCEs) flanking the rax coding region regulate expression of a rax promoter-GFP transgene in Xenopus tadpoles...
October 3, 2016: Genesis: the Journal of Genetics and Development
Qingxiang Zhou, Scott J Neal, Francesca Pignoni
A host of classical and molecular genetic tools make Drosophila a tremendous model for the dissection of gene activity. In particular, the FLP-FRT technique for mitotic recombination has greatly enhanced gene loss-of-function analysis. This technique efficiently induces formation of homozygous mutant clones in tissues of heterozygous organisms. However, the dependence of the FLP-FRT method on cell division, and other constraints, also impose limits on its effectiveness. We describe here the generation and testing of tools for Mutant Analysis by Rescue Gene Excision (MARGE), an approach whereby mutant cells are formed by loss of a rescue transgene in a homozygous mutant organism...
October 3, 2016: Genesis: the Journal of Genetics and Development
Kazuhito Sakamoto, Barbara L Wehde, Patrick D Rädler, Aleata A Triplett, Kay-Uwe Wagner
The biological functions of the Janus kinase 1 (JAK1) are suggested to be pleiotropic since this signal transducer is ubiquitously expressed and coupled to a variety of cytokine receptors. Consequently, mice that are deficient in this tyrosine kinase were reported to die shortly after birth. To facilitate studies that address the biological and molecular functions of JAK1 during postnatal development, we performed gene targeting in embryonic stem cells and generated a Cre/lox-based conditional knockout mouse model...
September 27, 2016: Genesis: the Journal of Genetics and Development
Ernesto Lujan, Douglas J Bornemann, Carmen Rottig, Brian A Bayless, Hugo Stocker, Ernst Hafen, Kavita Arora, Rahul Warrior
The FLP/FRT system permits rapid phenotypic screening of homozygous lethal mutations in the context of a viable mosaic fly. Combining this system with ovo(D) dominant female-sterile transgenes enables efficient production of embryos derived from mutant germline clones lacking maternal contribution from a gene of interest. Two distinct sets of FRT chromosomes, carrying either the mini-white (w (+ mW.hs) ), or rosy (ry(+) ) and neomycin (neo(R) ) transgenes are in common use. Parallel ovo(D) lines were developed using w (+ mW...
September 16, 2016: Genesis: the Journal of Genetics and Development
Shenyue Qin, Mayur Madhavan, Ronald R Waclaw, Masato Nakafuku, Kenneth Campbell
In this study, we generated a transgenic mouse line driving Cre and EGFP expression with two putative cis-regulatory elements (CRMs) (i.e. hs687 and hs678) upstream of the homeobox gene Gsx2 (formerly Gsh2), a critical gene for establishing lateral ganglionic eminence (LGE) identity. The combination of these two CRMs drives transgene expression within the endogenous Gsx2 expression domains along the anterior-posterior neuraxis. By crossing this transgenic line with the Rosa(tdTomato) (Ai14) reporter mouse line, we observed a unique recombination pattern in the lateral ventral telencephalon, namely the LGE and the dorsal half of the medial GE (MGE), but not in the septum...
September 12, 2016: Genesis: the Journal of Genetics and Development
Hidemi Misawa, Daijiro Inomata, Miseri Kikuchi, Sae Maruyama, Yasuhiro Moriwaki, Takashi Okuda, Nobuyuki Nukina, Tomoyuki Yamanaka
VAChT-Cre.Fast and VAChT-Cre.Slow mice selectively express Cre recombinase in approximately one half of post-natal somatic motor neurons. The mouse lines have been used in various studies with selective genetic modifications in adult motor neurons. In the present study, we crossed VAChT-Cre lines with a reporter line, CAG-Syp/tdTomato, in which synaptophysin-tdTomato fusion proteins are efficiently sorted to axon terminals, making it possible to label both cell bodies and axon terminals of motor neurons. In the mice, Syp/tdTomato fluorescence preferentially co-localized with osteopontin, a recently discovered motor neuron marker for slow-twitch fatigue-resistant (S) and fast-twitch fatigue-resistant (FR) types...
September 6, 2016: Genesis: the Journal of Genetics and Development
Sang-Chul Nam
Cell polarity genes have important functions in photoreceptor morphogenesis. Based on recent discovery of stabilized microtubule cytoskeleton in developing photoreceptors and its role in photoreceptor cell polarity, microtubule associated proteins might have important roles in controlling cell polarity proteins' localizations in developing photoreceptors. Here, Tau, a microtubule associated protein, was analyzed to find its potential role in photoreceptor cell polarity. Tau co-localizes with acetylated/stabilized microtubules in developing pupal photoreceptors...
August 31, 2016: Genesis: the Journal of Genetics and Development
Dongwang Zheng, Xiaoyan Yang, Donglai Sheng, Dongliang Yu, Guoqing Liang, Luming Guo, Mei Xu, Xu Hu, Daqiang He, Yang Yang, Yuying Wang
Pou4f2 acts as a key node in the comprehensive and step-wise gene regulatory network (GRN) and regulates the development of retinal ganglion cells (RGCs). Accordingly, deletion of Pou4f2 results in RGC axon defects and apoptosis. To investigate the GRN involved in RGC regeneration, we generated a mouse line with a POU4F2-green fluorescent protein (GFP) fusion protein expressed in RGCs. Co-localization of POU4F2 and GFP in the retina and brain of Pou4f2-GFP/+ heterozygote mice was confirmed using immunofluorescence analysis...
August 17, 2016: Genesis: the Journal of Genetics and Development
Edwin Chaharbakhshi, Jennifer C Jemc
Broad-complex, Tramtrack, and Bric-à-brac/poxvirus and zinc finger (BTB/POZ) family proteins are a diverse family of proteins that are characterized by the presence of a common protein-protein interaction domain, known as the BTB domain. BTB proteins have been identified in poxviruses and many eukaryotes, and have diverse functions, ranging from transcriptional regulation and chromatin remodeling to protein degradation and cytoskeletal regulation. Specificity of function is determined in part by additional domains present in BTB family proteins, as well as by interaction partners...
August 13, 2016: Genesis: the Journal of Genetics and Development
Ming Yu, Salma Al-Dallal, Latifa Al-Haj, Shiraj Panjwani, Akina S McCartney, Sarah M Edwards, Pooja Manjunath, Catherine Walker, Alexander Awgulewitsch, Kathryn E Hentges
The mouse zinc-finger gene Zfp521 (also known as ecotropic viral insertion site 3; Evi3; and ZNF521 in humans) has been identified as a B-cell proto-oncogene, causing leukaemia in mice following retroviral insertions in its promoter region that drive Zfp521 over-expression. Furthermore, ZNF521 is expressed in human haematopoietic cells, and translocations between ZNF521 and PAX5 are associated with paediatric acute lymphoblastic leukaemia. However, the regulatory factors that control Zfp521 expression directly have not been characterised...
August 10, 2016: Genesis: the Journal of Genetics and Development
Reza K Oqani, Tao Lin, Jae Eun Lee, So Yeon Kim, Soo Jin Sa, Je Seok Woo, Dong Il Jin
Positive transcription elongation factor b (P-TEFb) is an RNA polymerase II kinase that phosphorylates Ser2 of the carboxyl-terminal domain and promotes the elongation phase of transcription. Despite the fact that P-TEFb has role in many cellular processes, the role of this kinase complex remains to be understood in early developmental events. In this study, using immunocytochemical analyses, we find that the P-TEFb components, Cyclin T1, CDK9, and its T-loop phosphorylated form, are localized to nuclear speckles, as well as in nucleoli in mouse germinal vesicle oocytes...
September 2016: Genesis: the Journal of Genetics and Development
Ravindra B Chalamalasetty, Rieko Ajima, Robert J Garriock, Mark W Kennedy, Lino Tessarollo, Terry P Yamaguchi
Wnt/β-catenin signals are important regulators of embryonic and adult stem cell self-renewal and differentiation and play causative roles in tumorigenesis. Purified recombinant Wnt3a protein, or Wnt3a-conditioned culture medium, has been widely used to study canonical Wnt signaling in vitro or ex vivo. To study the role of Wnt3a in embryogenesis and cancer models, we developed a Cre recombinase activatable Rosa26(Wnt3a) allele, in which a Wnt3a cDNA was inserted into the Rosa26 locus to allow for conditional, spatiotemporally defined expression of Wnt3a ligand for gain-of-function (GOF) studies in mice...
September 2016: Genesis: the Journal of Genetics and Development
Amanda N Marra, Yue Li, Rebecca A Wingert
Cilia arose early during eukaryotic evolution, and their structural components are highly conserved from the simplest protists to complex metazoan species. In recent years, the role of cilia in the ontogeny of vertebrate organs has received increasing attention due to a staggering correlation between human disease and dysfunctional cilia. In particular, the presence of cilia in both the developing and mature kidney has become a deep area of research due to ciliopathies common to the kidney, such as polycystic kidney disease (PKD)...
September 2016: Genesis: the Journal of Genetics and Development
Jifan Feng, Junjun Jing, Pedro A Sanchez-Lara, Moiz S Bootwalla, Jonathan Buckley, Nancy Wu, Youzhen Yan, Yang Chai
Pax9 encodes a paired-box homeodomain (Pax) transcription factor and is critical for the development of multiple organs. Using CrispR/Cas9-mediated homologous directed repair (HDR), we generated a new Pax9-CreER knock-in mouse line in which the CreER(T2) fusion protein is produced after synthesis of endogenous Pax9 protein. We found that tdTomato reporter expression in Pax9-CreER;tdTomato reporter mice is detectable in a similar pattern to the endogenous Pax9 expression, faithfully recapitulating the Pax9 expression domains throughout the embryo and in the adult mouse...
September 2016: Genesis: the Journal of Genetics and Development
Takamasa Mizoguchi, Koichi Kawakami, Motoyuki Itoh
Actin filaments and microtubules are principal components of the cytoskeleton that regulate the basic cellular phenomena underlying many fundamental cellular processes. Therefore, analyzing their dynamics in living cells is important for understanding cellular events more precisely. In this article, we report two novel transgenic zebrafish lines expressing red fluorescent proteins tagged with Lifeact or EB1 that interact with actin filaments and microtubule plus ends, respectively, under the control of the GAL4-UAS system...
September 2016: Genesis: the Journal of Genetics and Development
Nicholas W Plummer, Jacqueline de Marchena, Patricia Jensen
Engrailed 1 (En1) is a homeobox-containing transcription factor expressed during development in diverse tissues, including the embryonic midbrain and anterior hindbrain. To facilitate investigation of genetic and developmental heterogeneity among cells with a history of En1 expression, we have generated En1(Dre) , a knock-in allele expressing Dre recombinase. En1(Dre) can be used with existing Cre and Flp recombinase lines for genetic intersectional labeling, fate mapping, and functional manipulation of subpopulations of cells characterized by transient expression of En1...
August 2016: Genesis: the Journal of Genetics and Development
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