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https://www.readbyqxmd.com/read/27906050/the-sh3-and-cysteine-rich-domain-3-stac3-gene-is-important-to-growth-fiber-composition-and-calcium-release-from-the-sarcoplasmic-reticulum-in-postnatal-skeletal-muscle
#1
Xiaofei Cong, Jonathan Doering, Davi A G Mazala, Eva R Chin, Robert W Grange, Honglin Jiang
BACKGROUND: The SH3 and cysteine-rich domain 3 (Stac3) gene is specifically expressed in the skeletal muscle. Stac3 knockout mice die perinatally. In this study, we determined the potential role of Stac3 in postnatal skeletal muscle growth, fiber composition, and contraction by generating conditional Stac3 knockout mice. METHODS: We disrupted the Stac3 gene in 4-week-old male mice using the Flp-FRT and tamoxifen-inducible Cre-loxP systems. RESULTS: RT-qPCR and western blotting analyses of the limb muscles of target mice indicated that nearly all Stac3 mRNA and more than 70 % of STAC3 protein were deleted 4 weeks after tamoxifen injection...
April 11, 2016: Skeletal Muscle
https://www.readbyqxmd.com/read/27899315/injury-stimulated-sonic-hedgehog-expression-in-microglia-contributes-to-neuroinflammatory-response-in-the-mptp-model-of-parkinson-s-disease
#2
Jung Hwi Lee, Young Cheul Chung, Eugene Bok, Hankyu Lee, Sue Hee Huh, Ji Eun Lee, Byung Kwan Jin, Hyuk Wan Ko
Parkinson's disease (PD) is a progressive neurodegenerative disorder in which dopamine (DA) neurons in the substantia nigra pars compacta (SNpc) region are selectively destroyed. Sonic hedgehog (Shh) has been well known to play a key role in a variety of processes such as embryogenesis, cell proliferation and protection, and tissue repair during inflammation. However, the evidences for the innate role of Shh in adult brain injury are presently lacking and studies have been needed to unveil the importance of Shh in the process of neurodegeneration...
November 26, 2016: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/27895309/hepatocyte-specific-expression-of-an-oncogenic-variant-of-%C3%AE-catenin-results-in-cholestatic-liver-disease
#3
Ursula J Lemberger, Claudia Fuchs, Matthias Karer, Stefanie Haas, Tatjana Stojakovic, Christian Schöfer, Hanns-Ulrich Marschall, Fritz Wrba, Makoto M Taketo, Gerda Egger, Michael Trauner, Christoph H Österreicher
BACKGROUND: The Wnt/β-catenin signaling pathway plays a crucial role in embryonic development, tissue homeostasis, wound healing and malignant transformation in different organs including the liver. The consequences of continuous β-catenin signaling in hepatocytes remain elusive. RESULTS: Livers of Ctnnb1CA hep mice were characterized by disturbed liver architecture, proliferating cholangiocytes and biliary type of fibrosis. Serum ALT and bile acid levels were significantly increased in Ctnnb1CA hep mice...
November 23, 2016: Oncotarget
https://www.readbyqxmd.com/read/27880796/foxn1-cre-expression-in-the-male-germline
#4
Jianjun Shi, Irina Getun, Bivian Torres, Howard T Petrie
Foxn1 (forkhead box N1), also known as the nude gene or winged-helix nude (Whn), is a forkhead transcription factor thought to be restricted to keratinocytes in the skin and thymus. Consistent with this tissue distribution, spontaneous or targeted mutation of Foxn1 results in the absence of both hair and a thymus. Genetic manipulation of the Foxn1 locus thus represents a powerful tool for tissue specific gene control in the skin and thymus, and tools such as Cre recombinase under control of the Foxn1 locus are widely used for this purpose...
2016: PloS One
https://www.readbyqxmd.com/read/27878291/adenovirus-mediated-truncated-bid-overexpression-induced-by-the-cre-loxp-system-promotes-the-cell-apoptosis-of-cd133-ovarian-cancer-stem-cells
#5
Qifang Long, Ru Yang, Weixian Lu, Weipei Zhu, Jundong Zhou, Cui Zheng, Dongmei Zhou, Ling Yu, Jinchang Wu
Cancer stem cells are a small subset of cancer cells that contribute to cancer progression, metastasis, chemoresistance and recurrence. CD133-positive (CD133+) ovarian cancer cells have been identified as ovarian cancer stem cells. Adenovirus-mediated gene therapy is an innovative therapeutic method for cancer treatment. In the present study, we aimed to develop a new gene therapy to specifically eliminate CD133+ ovarian cancer stem cells by targeting CD133. We used the Cre/LoxP system to augment the selective expression of the truncated Bid (tBid) gene as suicide gene therapy in CD133+ ovarian cancer stem cells...
November 21, 2016: Oncology Reports
https://www.readbyqxmd.com/read/27870307/abrogation-of-gap-junctional-communication-in-es-cells-results-in-a-disruption-of-primitive-endoderm-formation-in-embryoid-bodies
#6
Philipp Wörsdörfer, Felicitas Bosen, Martina Gebhardt, Nicole Russ, Katrin Zimmermann, David Komla Kessie, Thileepan Sekaran, Angela Egert, Süleyman Ergün, Hubert Schorle, Alexander Pfeifer, Frank Edenhofer, Klaus Willecke
Gap junctional intercellular communication (GJIC) has been suggested to be involved in early embryonic development but the actual functional role remained elusive. Connexin (Cx) 43 and Cx45 are co-expressed in embryonic stem (ES) cells, form gap junctions and are considered to exhibit adhesive function and/or to contribute to the establishment of defined communication compartments. Here we describe the generation of Cx43/Cx45-double deficient mouse ES cells to achieve almost complete breakdown of GJIC. Cre-loxP induced deletion of both, Cx43 and Cx45, results in a block of differentiation in embryoid bodies without affecting pluripotency marker expression and proliferation in ES cells...
November 21, 2016: Stem Cells
https://www.readbyqxmd.com/read/27868059/intramyocardial-injection-of-sirnas-can-efficiently-establish-myocardial-tissue-specific-renalase-knockdown-mouse-model
#7
Kun Huang, Ju Liu, Hui Zhang, Jiliang Wang, Huili Li
Ischaemia/reperfusion (I/R) injury will cause additional death of cardiomyocytes in ischaemic heart disease. Recent studies revealed that renalase was involved in the I/R injury. So, the myocardial tissue-specific knockdown mouse models were needed for the investigations of renalase. To establish the mouse models, intramyocardial injection of siRNAs targeting renalase was performed in mice. The wild distribution and high transfection efficiency of the siRNAs were approved. And the renalase expression was efficiently suppressed in myocardial tissue...
2016: BioMed Research International
https://www.readbyqxmd.com/read/27859428/low-p16-ink4a-expression-in-early-passage-human-prostate-basal-epithelial-cells-enables-immortalization-by-telomerase-expression-alone
#8
Mindy Kim Graham, Lorenzo Principessa, Lizamma Antony, Alan K Meeker, John T Isaacs
BACKGROUND: There are two principal senescence barriers that must be overcome to successfully immortalize primary human epithelial cells in culture, stress-induced senescence, and replicative senescence. The p16(INK4a) /retinoblastoma protein (p16/Rb) pathway mediates stress-induced senescence, and is generally upregulated by primary epithelial cells in response to the artificial conditions from tissue culture. Replicative senescence is associated with telomere loss. Following each round of cell division, telomeres progressively shorten...
November 8, 2016: Prostate
https://www.readbyqxmd.com/read/27856232/improved-recombinant-antibody-production-by-cho-cells-using-a-production-enhancer-dna-element-with-repeated-transgene-integration-at-a-predetermined-chromosomal-site
#9
Yoshinori Kawabe, Takanori Inao, Shodai Komatsu, Guan Huang, Akira Ito, Takeshi Omasa, Masamichi Kamihira
Chinese hamster ovary (CHO) cells are one of the most useful host cell lines for the production of biopharmaceutical proteins. Although a series of production processes have been refined to improve protein productivity and cost performance, establishing producer cells is still time-consuming and labor-intensive. Recombinase-mediated site-specific gene integration into a predetermined chromosomal locus may enable predictable protein expression, reducing the laborious process of cell screening. We previously developed an accumulative site-specific gene integration system (AGIS) using Cre recombinase and mutated loxP sites for transgene integration and amplification in the CHO cell genome...
November 14, 2016: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/27848935/distinct-roles-of-creb-within-the-ventral-and-dorsal-hippocampus-in-mediating-nicotine-withdrawal-phenotypes
#10
Miranda Fisher, Rachel LeMalefant, Luyi Zhou, Gavin Huang, Jill Turner
Addiction to nicotine and the inability to quit smoking are influenced by genetic factors, emphasizing the importance of understanding how genes and drugs of abuse mechanistically impact each other. One well-characterized protein responsible for regulating both response to drugs and gene expression is the transcription factor CREB. Previous work indicates that hippocampal specific alterations in CREB signaling and synaptic plasticity may underlie certain nicotine withdrawal phenotypes. However, the structure of the hippocampus possesses dorsal and ventral sub-regions, each differing in behavioral, anatomic and gene expression characteristics...
November 16, 2016: Neuropsychopharmacology: Official Publication of the American College of Neuropsychopharmacology
https://www.readbyqxmd.com/read/27845447/novel-method-to-rescue-a-lethal-phenotype-through-integration-of-target-gene-onto-the-x-chromosome
#11
Kazuya Sakata, Kimi Araki, Hiroyasu Nakano, Takashi Nishina, Sachiko Komazawa-Sakon, Shin Murai, Grace E Lee, Daisuke Hashimoto, Chigure Suzuki, Yasuo Uchiyama, Kenji Notohara, Anna S Gukovskaya, Ilya Gukovsky, Ken-Ichi Yamamura, Hideo Baba, Masaki Ohmuraya
The loss-of-function mutations of serine protease inhibitor, Kazal type 1 (SPINK1) gene are associated with human chronic pancreatitis, but the underlying mechanisms remain unknown. We previously reported that mice lacking Spink3, the murine homologue of human SPINK1, die perinatally due to massive pancreatic acinar cell death, precluding investigation of the effects of SPINK1 deficiency. To circumvent perinatal lethality, we have developed a novel method to integrate human SPINK1 gene on the X chromosome using Cre-loxP technology and thus generated transgenic mice termed "X-SPINK1"...
November 15, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27837366/hippocampus-specific-deficiency-of-il-15r%C3%AE-contributes-to-greater-anxiety-like-behaviors-in-mice
#12
Linda Nguyen, Joseph Bohlen, Janelle Stricker, Ikttesh Chahal, Hanting Zhang, Emidio E Pistilli
A hippocampus-specific IL15RαKO mouse (hipIl15ra (fl/fl) /Cre(+)) was generated to test the hypothesis that the targeted deletion of interleukin-15 receptor alpha (IL-15Rα) in the hippocampus contributes to altered behavior, including greater levels of anxiety and ambulatory activity. Using Cre-loxP, exons 2 and 3 of the IL-15Rα gene were excised within the hippocampus, while normal expression was maintained within the rest of the brain. In the open field test (OFT), hipIl15ra (fl/fl) /Cre(+) spent a greater amount of time in the periphery and less time in the central portions of the chamber, and there was also a noticeable trend for decreased rearing activity; these behaviors are consistent with greater levels of anxiety-like behavior in these mice...
November 11, 2016: Metabolic Brain Disease
https://www.readbyqxmd.com/read/27836576/establishment-of-a-conditional-transgenic-mouse-model-recapitulating-eml4-alk-positive-human-non-small-cell-lung-cancer
#13
Kyoung Ho Pyo, Sun Min Lim, Hye Ryun Kim, Young Hoon Sung, Mi Ran Yun, Sung-Moo Kim, Hwan Kim, Han Na Kang, Ji Min Lee, Sang Gyun Kim, Chae Won Park, Hyun Chang, Hyo Sup Shim, Han-Woong Lee, Byoung Chul Cho
BACKGROUND: ALK fusion is a distinct molecular subclassification of non-small-cell lung cancer (NSCLC) that is targeted by ALK inhibitors. We established a transgenic mouse model that expresses tumors highly resembling human NSCLC harboring EML-ALK fusion. We aimed to test EML4-ALK transgenic mouse model as a platform for assessing efficacy of ALK inhibitor and examining mechanisms of acquired resistance to ALK inhibitor. MATERIALS AND METHODS: Transgenic mouse lines harboring LoxP-STOP-LoxP- FLAG tagged human EML4-ALK (variant 1) transgene was established using C57BL/6N mice...
November 8, 2016: Journal of Thoracic Oncology
https://www.readbyqxmd.com/read/27834664/%C3%AE-sma-expressing-perivascular-cells-represent-dental-pulp-progenitors-in-vivo
#14
I Vidovic, A Banerjee, R Fatahi, B G Matthews, N A Dyment, I Kalajzic, M Mina
The goal of this study was to examine the contribution of perivascular cells to odontoblasts during the development, growth, and repair of dentin using mouse molars as a model. We used an inducible, Cre-loxP in vivo fate-mapping approach to examine the contributions of the descendants of cells expressing the αSMA-CreERT2 transgene to the odontoblast lineage. In vivo lineage-tracing experiments in molars showed the contribution of αSMA-tdTomato(+) cells to a small number of newly formed odontoblasts during primary dentinogenesis...
November 10, 2016: Journal of Dental Research
https://www.readbyqxmd.com/read/27829235/carf-activates-beta-catenin-tcf-signaling-in-the-hepatocellular-carcinoma
#15
Xin Fan, Xiaoyan Ma, Lei Cui, Shengchun Dang, Jianguo Qu, Jianxin Zhang, Xuqing Wang, Zhengfa Mao
Overactivation of Ras signaling is very common in the hepatocellular carcinoma (HCC) due to its constitutive active mutation, which makes it a big challenge to target Ras signaling. Therefore, identifying effectors downstream of Ras signaling would benefit the development of novel therapeutic strategies. In this study, it was found that the expression of CARF (collaborate of ARF) was induced by oncogenic RasV12. The expression of CARF was up-regulated in both HCC mouse model (Alb-Cre; P53f/f; Loxp-Stop-Loxp-RasG12D) and human HCC clinical samples...
November 5, 2016: Oncotarget
https://www.readbyqxmd.com/read/27826231/mice-lacking-brinp2-or-brinp3-or-both-exhibit-behaviors-consistent-with-neurodevelopmental-disorders
#16
Susan R Berkowicz, Travis J Featherby, James C Whisstock, Phillip I Bird
Background:Brinps 1-3, and Astrotactins (Astn) 1 and 2, are members of the Membrane Attack Complex/Perforin (MACPF) superfamily that are predominantly expressed in the mammalian brain during development. Genetic variation at the human BRINP2/ASTN1 and BRINP1/ASTN2 loci has been implicated in neurodevelopmental disorders. We, and others, have previously shown that Brinp1(-/-) mice exhibit behavior reminiscent of autism spectrum disorder (ASD) and attention deficit hyperactivity disorder (ADHD). Method: We created Brinp2(-/-) mice and Brinp3(-/-) mice via the Cre-mediated LoxP system to investigate the effect of gene deletion on anatomy and behavior...
2016: Frontiers in Behavioral Neuroscience
https://www.readbyqxmd.com/read/27825338/sequential-and-counter-selectable-cassettes-for-fission-yeast
#17
Hanna Amelina, Vera Moiseeva, Laura Catharine Collopy, Siân Rosanna Pearson, Christine Anne Armstrong, Kazunori Tomita
BACKGROUND: Fission yeast is one of the most commonly used model organisms for studying genetics. For selection of desirable genotypes, antibiotic resistance cassettes are widely integrated into the genome near genes of interest. In yeasts, this is achieved by PCR amplification of the cassette flanked by short homology sequences, which can be incorporated by homology directed repair. However, the currently available cassettes all share the same tef promoter and terminator sequences. It can therefore be challenging to perform multiple genetic modifications by PCR-based targeting, as existing resistance cassettes in strains can be favored for recombination due to shared homology between the cassettes...
November 8, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27812322/tamoxifen-in-the-mouse-brain-implications-for-fate-mapping-studies-using-the-tamoxifen-inducible-cre-loxp-system
#18
Martin Valny, Pavel Honsa, Denisa Kirdajova, Zdenek Kamenik, Miroslava Anderova
The tamoxifen-inducible Cre-loxP system is widely used to overcome gene targeting pre-adult lethality, to modify a specific cell population at desired time-points, and to visualize and trace cells in fate-mapping studies. In this study we focused on tamoxifen degradation kinetics, because for all genetic fate-mapping studies, the period during which tamoxifen or its metabolites remain active in the CNS, is essential. Additionally, we aimed to define the tamoxifen administration scheme, enabling the maximal recombination rate together with minimal animal mortality...
2016: Frontiers in Cellular Neuroscience
https://www.readbyqxmd.com/read/27796960/lineage-tracing-of-mammary-stem-and-progenitor-cells
#19
Anoeska A A van de Moosdijk, Nai Yang Fu, Anne C Rios, Jane E Visvader, Renée van Amerongen
Lineage tracing analysis allows mammary epithelial cells to be tracked in their natural environment, thereby revealing cell fate and proliferation choices in the intact tissue. This technique is particularly informative for studying how stem cells build and maintain the mammary epithelium during development and pregnancy. Here we describe two experimental systems based on Cre/loxP technology (Cre(ERT2)/loxP and rtTA/tetO-Cre/loxP), which allow the inducible, permanent labeling of mammary epithelial cells following the administration of either tamoxifen or doxycycline...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27793816/the-transcription-factor-gli2-as-a-downstream-mediator-of-transforming-growth-factor-%C3%AE-induced-fibroblast-activation-in-ssc
#20
Ruifang Liang, Barbora Šumová, Cinzia Cordazzo, Tatjana Mallano, Yun Zhang, Thomas Wohlfahrt, Clara Dees, Andreas Ramming, Dorota Krasowska, Małgorzata Michalska-Jakubus, Oliver Distler, Georg Schett, Ladislav Šenolt, Jörg H W Distler
OBJECTIVES: Hedgehog signalling plays a critical role during the pathogenesis of fibrosis in systemic sclerosis (SSc). Besides canonical hedgehog signalling with smoothened (SMO)-dependent activation of GLI transcription factors, GLI can be activated independently of classical hedgehog ligands and receptors (so-called non-canonical pathways). Here, we aimed to evaluate the role of non-canonical hedgehog signalling in SSc and to test the efficacy of direct GLI inhibitors that target simultaneously canonical and non-canonical hedgehog pathways...
October 28, 2016: Annals of the Rheumatic Diseases
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