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Cre/loxp

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https://www.readbyqxmd.com/read/29774106/hematopoietic-restricted-ptpn11e76k-reveals-indolent-mpn-progression-in-mice
#1
Stefan P Tarnawsky, Wen-Mei Yu, Cheng-Kui Qu, Rebecca J Chan, Mervin C Yoder
Juvenile Myelomonocytic Leukemia (JMML) is a pediatric myeloproliferative neoplasm (MPN) that has a poor prognosis. Somatic mutations in Ptpn11 are the most frequent cause of JMML and they commonly occur in utero . Animal models of mutant Ptpn11 have probed the signaling pathways that contribute to JMML. However, existing models may inappropriately exacerbate MPN features by relying on non-hematopoietic-restricted Cre-loxP strains or transplantations into irradiated recipients. In this study we generate hematopoietic-restricted models of Ptpn11E76K-mediated disease using Csf1r-MCM and Flt3Cre...
April 24, 2018: Oncotarget
https://www.readbyqxmd.com/read/29769309/etoposide-induced-protein-2-4-functions-as-a-regulator-of-the-calcium-atpase-and-protects-pancreatic-%C3%AE-cell-survival
#2
Lin Yuan, Huiyu Wang, Qi Liu, Zhe Wang, Mingshu Zhang, Yan Zhao, Kuo Liang, Liangyi Chen, Tao Xu, Pingyong Xu
Calcium homeostasis is essential for maintaining the viability and function of pancreatic β cells and plays a key role in preventing the development of diabetes. Decreased levels of ATPase sarco-plasmic/endoplasmic reticulum Ca2+ -transporting 2 (ATP2a2), the main calcium pump in β cells, are often found in individuals with diabetes and in diabetic animal models. However, the regulators of ATP2a2 and the molecular mechanisms responsible for controlling ATP2a2 activity remain unclear. Etoposide-induced protein 2...
May 16, 2018: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/29764841/genetic-targeting-of-organ-specific-blood-vessels
#3
Wenjuan Pu, Lingjuan He, Ximeng Han, Xueying Tian, Yan Li, Hui Zhang, Qiaozhen Liu, Xiuzhen Huang, Libo Zhang, Qing-Dong Wang, Zhenyang Yu, Xiao Yang, Nicola Smart, Bin Zhou
<u>Rationale:</u> Organs of the body require vascular networks to supply oxygen and nutrients and maintain physiological function. The blood vessels of different organs are structurally and functionally heterogeneous in nature. To more precisely dissect their distinct in vivo function in individual organs, without potential interference from off-site targets, it is necessary to genetically target them in an organ-specific manner. <u>Objective:</u> To generate a genetic system that targets vascular endothelial cells in an organ- or tissue- specific manner and to exemplify the potential application of intersectional genetics for precise, target-specific gene manipulation in vivo...
May 15, 2018: Circulation Research
https://www.readbyqxmd.com/read/29761406/cre-driver-mice-targeting-macrophages
#4
Jiayuan Shi, Li Hua, Danielle Harmer, Peishan Li, Guangwen Ren
The Cre/loxP system is a widely applied technology for site-specific genetic manipulation in mice. This system allows for deletion of the genes of interest in specific cells, tissues, and whole organism to generate a diversity of conditional knockout mouse strains. Additionally, the Cre/loxP system is useful for development of cell- and tissue-specific reporter mice for lineage tracing, and cell-specific conditional depletion models in mice. Recently, the Cre/loxP technique was extensively adopted to characterize the monocyte/macrophage biology in mouse models...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29743652/adeno-associated-virus-2-9-delivery-of-cre-recombinase-in-mouse-primary-afferents
#5
Khaled Abdallah, Francis Nadeau, Francis Bergeron, Sylvie Blouin, Véronique Blais, Kelly M Bradbury, Christine L Lavoie, Jean-Luc Parent, Louis Gendron
Genetically-modified animal models have significantly increased our understanding of the complex central nervous system circuits. Among these models, inducible transgenic mice whose specific gene expression can be modulated through a Cre recombinase/LoxP system are useful to study the role of specific peptides and proteins in a given population of cells. In the present study, we describe an efficient approach to selectively deliver a Cre-GFP to dorsal root ganglia (DRG) neurons. First, mice of different ages were injected in both hindpaws with a recombinant adeno-associated virus (rAAV2/9-CBA-Cre-GFP)...
May 9, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29743638/screen-and-verification-for-transgene-integration-sites-in-pigs
#6
Linyuan Ma, Yuzhe Wang, Haitao Wang, Yiqing Hu, Jingyao Chen, Tan Tan, Man Hu, Xiaojuan Liu, Ran Zhang, Yiming Xing, Yiqiang Zhao, Xiaoxiang Hu, Ning Li
Efficient transgene expression in recipient cells constitutes the primary step in gene therapy. However, random integration in host genome comprises too many uncertainties. Our study presents a strategy combining bioinformatics and functional verification to find transgene integration sites in pig genome. Using an in silico approach, we screen out two candidate sites, namely, Pifs302 and Pifs501, located in actively transcribed intergenic regions with low nucleosome formation potential and without potential non-coding RNAs...
May 9, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29743295/defining-the-tissue-specific-roles-of-igf1-igf1r-in-regulating-metabolism-using-the-cre-loxp-system
#7
Rhonda D Kineman, Mercedes Del Rio-Moreno, André Sarmento-Cabral
It is clear that IGF1 is important in supporting growth and regulating metabolism. The IGF1 found in the circulation is primarily produced by the liver hepatocytes, but healthy mature hepatocytes do not express appreciable levels of the IGF1 receptor (IGF1R). Therefore, the metabolic actions of IGF1 are thought to be mediated via extra-hepatocyte actions. Given the structural and functional homology between IGF1/IGF1R and insulin receptor (INSR) signaling, and the fact that IGF1, IGF1R and INSR are expressed in most tissues of the body, it is difficult to separate out the tissue-specific contributions of IGF1/IGF1R in maintaining whole body metabolic function...
May 9, 2018: Journal of Molecular Endocrinology
https://www.readbyqxmd.com/read/29720668/central-role-of-the-proximal-tubular-%C3%AE-klotho-fgf-receptor-complex-in-fgf23-regulated-phosphate-and-vitamin-d-metabolism
#8
Ai Takeshita, Kazuki Kawakami, Kenryo Furushima, Masayasu Miyajima, Kazushige Sakaguchi
Fibroblast growth factor 23 (FGF23) plays critical roles in phosphate handling and vitamin D metabolism in the kidney. However, the effector cells of FGF23 in the kidney remain unclear. αKlotho, a putative enzyme possessing β-glucuronidase activity and also a permissive co-receptor for FGF23 to bind to FGF receptors (FGFRs), is expressed most abundantly in distal convoluted tubules, whereas it is expressed modestly in proximal tubules. Key molecular players of phosphate homeostasis and vitamin D-metabolizing enzymes are known to localize in proximal tubules...
May 2, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29720086/click-one-step-generation-of-conditional-knockout-mice
#9
Yoshiki Miyasaka, Yoshihiro Uno, Kazuto Yoshimi, Yayoi Kunihiro, Takuji Yoshimura, Tomohiro Tanaka, Harumi Ishikubo, Yuichi Hiraoka, Norihiko Takemoto, Takao Tanaka, Yoshihiro Ooguchi, Paul Skehel, Tomomi Aida, Junji Takeda, Tomoji Mashimo
BACKGROUND: CRISPR/Cas9 enables the targeting of genes in zygotes; however, efficient approaches to create loxP-flanked (floxed) alleles remain elusive. RESULTS: Here, we show that the electroporation of Cas9, two gRNAs, and long single-stranded DNA (lssDNA) into zygotes, termed CLICK (CRISPR with lssDNA inducing conditional knockout alleles), enables the quick generation of floxed alleles in mice and rats. CONCLUSIONS: The high efficiency of CLICK provides homozygous knock-ins in oocytes carrying tissue-specific Cre, which allows the one-step generation of conditional knockouts in founder (F0) mice...
May 2, 2018: BMC Genomics
https://www.readbyqxmd.com/read/29701326/cre-mediated-transgene-integration-in-chinese-hamster-ovary-cells-using-minicircle-dna-vectors
#10
Xue Wang, Yoshinori Kawabe, Takeshi Hada, Akira Ito, Masamichi Kamihira
Bacterial backbone sequences of conventional plasmid vectors have been reported to exhibit negative effects on transgene expression in mammalian cells, such as cytotoxicity and gene silencing. Minicircle DNA vectors can be employed to overcome these issues and to improve the transfection efficiency because of their smaller size. In this study, transgenes were integrated into the hypoxanthine phosphoribosyltransferase (hprt) locus of Chinese hamster ovary (CHO) cells by the Cre-loxP system using minicircle DNA vectors as transgene donors...
April 27, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29698062/targeted-deletion-of-t-cell-s1p-receptor-1-ameliorates-cardiac-fibrosis-in-streptozotocin-induced-diabetic-mice
#11
Chowdhury S Abdullah, Zhu-Qiu Jin
Infiltration of T cells is associated with patients who have diabetes at an increased risk of heart attack. T-cell sphingosine 1-phosphate receptor 1 (S1P1 )-mediated signaling directs T lymphocyte trafficking. Effects of T-cell S1P1 activation on cardiac fibrosis in a murine diabetic model remain to be explored. For this purpose, conditional T-cell S1P1 knockout (TS1P1 KO) mice generated by crossing S1pr1 loxP/loxP mice with Lck-Cre mice were used in a model of streptozotocin-induced diabetic cardiomyopathy...
April 26, 2018: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
https://www.readbyqxmd.com/read/29695773/stromal-interaction-molecule-1-modulates-blood-pressure-via-no-production-in-vascular-endothelial-cells
#12
Mitsuhiro Nishimoto, Risuke Mizuno, Toshiro Fujita, Masashi Isshiki
In vascular endothelial cells, store-operated calcium entry (SOCE) activates endothelial NO synthase (eNOS) and regulates nitric oxide (NO) production as well as flow-dependent mechanical stimuli. Stromal interaction molecule 1, or STIM1, was recently identified to be essential for SOCE, acting as a calcium sensor for intracellular calcium stores. However, how STIM1 affects endothelial function and blood pressure (BP) remains unclear. We generated STIM1 fl/fl mice and vascular endothelial cell-specific STIM1 knockout mice using the Cre-loxP system, and conducted experiments using these mice to clarify the physiological role of STIM1 in vascular endothelial function and BP as follows: (1) SOCE was analyzed in isolated aortic endothelial cells by calcium add-back with fluorescent Ca2+ indicators...
April 25, 2018: Hypertension Research: Official Journal of the Japanese Society of Hypertension
https://www.readbyqxmd.com/read/29683207/the-wmn1-enhancer-region-in-intron-1-is-required-for-expression-of-human-plp1
#13
Hamdan Hamdan, Pankaj Patyal, Neriman T Kockara, Patricia A Wight
The myelin proteolipid protein gene (PLP1) encodes the most abundant protein present in myelin from the central nervous system (CNS). Its expression must be tightly controlled as evidenced by mutations that alter PLP1 dosage; both overexpression (elevated PLP1 copy number) and lack thereof (PLP1 deletion) result in X-linked genetic disorders in man. However, not much is known about the mechanisms that govern expression of the human gene. To address this, transgenic mice were generated which utilize human PLP1 (hPLP1) sequences (proximal 6...
April 23, 2018: Glia
https://www.readbyqxmd.com/read/29673028/cre-driver-lines-used-for-genetic-fate-mapping-of-neural-crest-cells-in-the-mouse-an-overview
#14
REVIEW
Julien Debbache, Vadims Parfejevs, Lukas Sommer
The neural crest is one of the embryonic structures with the broadest developmental potential in vertebrates. Morphologically, neural crest cells emerge during neurulation in the dorsal folds of the neural tube before undergoing an epithelial-to-mesenchymal transition (EMT), delaminating from the neural tube, and migrating to multiple sites in the growing embryo. Neural crest cells generate cell types as diverse as peripheral neurons and glia, melanocytes, and so-called mesectodermal derivatives that include craniofacial bone and cartilage and smooth muscle cells in cardiovascular structures...
April 19, 2018: Genesis: the Journal of Genetics and Development
https://www.readbyqxmd.com/read/29671710/functionally-complete-excision-of-conditional-alleles-in-the-mouse-suprachiasmatic-nucleus-by-vgat-ires-cre
#15
David R Weaver, Vincent van der Vinne, E Lela Giannaris, Thomas J Vajtay, Kristopher L Holloway, Christelle Anaclet
Mice with targeted gene disruption have provided important information about the molecular mechanisms of circadian clock function. A full understanding of the roles of circadian-relevant genes requires manipulation of their expression in a tissue-specific manner, ideally including manipulation with high efficiency within the suprachiasmatic nuclei (SCN). To date, conditional manipulation of genes within the SCN has been difficult. In a previously developed mouse line, Cre recombinase was inserted into the vesicular GABA transporter (Vgat) locus...
April 2018: Journal of Biological Rhythms
https://www.readbyqxmd.com/read/29670283/genetic-identification-of-leptin-neural-circuits-in-energy-and-glucose-homeostases
#16
Jie Xu, Christopher L Bartolome, Cho Shing Low, Xinchi Yi, Cheng-Hao Chien, Peng Wang, Dong Kong
Leptin, a hormone produced in white adipose tissue, acts in the brain to communicate fuel status, suppress appetite following a meal, promote energy expenditure and maintain blood glucose stability1,2 . Dysregulation of leptin or its receptors (LEPR) results in severe obesity and diabetes3-5 . Although intensive studies on leptin have transformed obesity and diabetes research2,6 , clinical applications of the molecule are still limited 7 , at least in part owing to the complexity and our incomplete understanding of the underlying neural circuits...
April 18, 2018: Nature
https://www.readbyqxmd.com/read/29670094/loss-of-deptor-in-renal-tubules-protects-against-cisplatin-induced-acute-kidney-injury
#17
Caixia Wang, Huaiqian Dai, Zhi Xiong, Qiancheng Song, Zhipeng Zou, Mangmang Li, Jing Nie, Xiaochun Bai, Zhenguo Chen
DEP domain containing mTOR-interacting protein (DEPTOR) was originally identified as an in vivo dual inhibitor of mechanistic target of rapamycin (mTOR). It was recently reported to be involved in renal physiology and pathology in vitro; however, its detailed roles and mechanisms in vivo are completely unknown. We observed that DEPTOR expression in the kidney was markedly increased on day 3 after cisplatin treatment, at which time cell apoptosis peaked, implicating DEPTOR in cisplatin-induced acute kidney injury (AKI)...
April 18, 2018: Cell Death & Disease
https://www.readbyqxmd.com/read/29649366/fgf2-enhances-odontoblast-differentiation-by-%C3%AE-sma-progenitors-in-vivo
#18
I Vidovic-Zdrilic, K H Vining, A Vijaykumar, I Kalajzic, D J Mooney, M Mina
The goal of this study was to examine the effects of early and limited exposure of perivascular cells expressing α (αSMA) to fibroblast growth factor 2 (FGF2) in vivo. We performed in vivo fate mapping by inducible Cre-loxP and experimental pulp injury in molars to induce reparative dentinogenesis. Our results demonstrate that early delivery of exogenous FGF2 to exposed pulp led to proliferative expansion of αSMA-tdTomato+ cells and their accelerated differentiation into odontoblasts. In vivo lineage-tracing experiments showed that the calcified bridge/reparative dentin in FGF2-treated pulps were lined with an increased number of Dspp+ odontoblasts and devoid of BSP+ osteoblasts...
April 1, 2018: Journal of Dental Research
https://www.readbyqxmd.com/read/29644115/shp2-regulates-skeletal-cell-fate-by-modifying-sox9-expression-and-transcriptional-activity
#19
Chunlin Zuo, Lijun Wang, Raghavendra M Kamalesh, Margot E Bowen, Douglas C Moore, Mark S Dooner, Anthony M Reginato, Qian Wu, Christoph Schorl, Yueming Song, Matthew L Warman, Benjamin G Neel, Michael G Ehrlich, Wentian Yang
Chondrocytes and osteoblasts differentiate from a common mesenchymal precursor, the osteochondroprogenitor (OCP), and help build the vertebrate skeleton. The signaling pathways that control lineage commitment for OCPs are incompletely understood. We asked whether the ubiquitously expressed protein-tyrosine phosphatase SHP2 (encoded by Ptpn11 ) affects skeletal lineage commitment by conditionally deleting Ptpn11 in mouse limb and head mesenchyme using "Cre-loxP"-mediated gene excision. SHP2-deficient mice have increased cartilage mass and deficient ossification, suggesting that SHP2-deficient OCPs become chondrocytes and not osteoblasts...
2018: Bone Research
https://www.readbyqxmd.com/read/29626628/hepatocyte-specific-deletion-of-lass2-protects-against-diet-induced-hepatic-steatosis-and-insulin-resistance
#20
Shaohua Fan, Yanyan Wang, Cun Wang, Haojie Jin, Zheng Wu, Jun Lu, Zifeng Zhang, Chunhui Sun, Qun Shan, Dongmei Wu, Juan Zhuang, Ning Sheng, Ying Xie, Mengqiu Li, Bin Hu, Jingyuan Fang, Yuanlin Zheng, Wenxin Qin
Homo sapienslongevity assurance homolog 2 of yeast LAG1 (LASS2) is expressed mostly in human liver. Here, we explored roles of LASS2 in pathogenesis of hepatic steatosis. Hepatocyte-specific LASS2 knockout (LASS2-/- ) mice were generated using Cre-LoxP system. LASS2-/- and wild-type (WT) mice were fed with chow or high-fat diet (HFD). We found LASS2-/- mice were resistant to HFD-induced hepatic steatosis and insulin resistance. In HFD-fed mice, LASS2 deficiency significantly inhibited p38 MAPK and ERK1/ERK2 signaling in mouse liver...
April 4, 2018: Free Radical Biology & Medicine
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