Panoply™ Human GUCY2C Over-Expressing Stable Cell Line

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Panoply™ Human GUCY2C Over-expressing Stable Cell Line HEK293-GUCY2C Lot. No. (See product label) CELL LINE INFORMATION Cat.No. CSC-SC006776 Cell Line Description HEK293-GUCY2C cell line is clonally derived from HEK293 cell line, which has been transduced with a lentiviral vector encoding: 1) full length human GUCY2C under the CMV promoter, and 2) puromycin resistance gene under the control of PGK promoter. Overexpression of human GUCY2C has been quantified by qPCR. Background GUCY2C (guanylate cyclase 2C) is an enzyme that in humans is encoded by the GUCY2C gene. It is a transmembrane protein that functions as a receptor for endogenous peptides guanylin and uroguanylin, and the heat-stable E. coli enterotoxin. This protein activates the cystic fibrosis transmembrane conductance regulator. Mutations in this gene are associated with familial diarrhea (autosomal dominant) and meconium ileus (autosomal recessive). Host Cell HEK293 Gene Information Gene Name Guanylate cyclase 2C Official Symbol GUCY2C Species Homo sapiens (human) GeneID 2984 mRNA Reference NM_004963.3 Protein Reference NP_004954.2 Synonyms GC-C; STAR; DIAR6; GUC2C; MECIL; MUCIL SAFETY AND PACKAGING Mycoplasma Mycoplasma Status: Negative (MycoAlert Kit) Format One frozen vial Storage Liquid nitrogen Safety Considerations The following safety precautions should be observed. 1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum. 2. No eating, drinking or smoking while handling the stable line. 3. Wash hands after handling the stable line and before leaving the lab. 4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells. 5. All waste should be considered hazardous. 6. Dispose of all liquid waste after each experiment and treat with bleach. Creative Biogene. All rights reserved. 45-1 Ramsey Road Shirley, NY 11967, USA Tel: 631-626-9181 ·Fax:631-614-7828 E-mail: [email protected] http://www.creative-biogene.com/ Ship Dry ice Representative Data Figure 1. Cell morphology under white light Human GUCY2C mRNA Over-expression Level (2-ddCt) 70000 60000 50000 40000 30000 20000 10000 Fold increases in number in increases Fold 0 HEK293-hGUCY2C-Monoclonal-2 HEK293-hGUCY2C-Monoclonal-1 HEK293-Puro (Control) Samples Figure 2. Human GUCY2C qPCR data Creative Biogene. All rights reserved. 45-1 Ramsey Road Shirley, NY 11967, USA Tel: 631-626-9181 ·Fax:631-614-7828 E-mail: [email protected] http://www.creative-biogene.com/ .

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Meconium Ileus Caused by Mutations in GUCY2C, Encoding the CFTR-Activating Guanylate Cyclase 2C
View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector REPORT Meconium Ileus Caused by Mutations in GUCY2C, Encoding the CFTR-Activating Guanylate Cyclase 2C Hila Romi,1,6 Idan Cohen,1,6 Daniella Landau,2 Suliman Alkrinawi,2 Baruch Yerushalmi,2 Reli Hershkovitz,3 Nitza Newman-Heiman,2 Garry R. Cutting,4 Rivka Ofir,1 Sara Sivan,1 and Ohad S. Birk1,5,* Meconium ileus, intestinal obstruction in the newborn, is caused in most cases by CFTR mutations modulated by yet-unidentified modi- fier genes. We now show that in two unrelated consanguineous Bedouin kindreds, an autosomal-recessive phenotype of meconium ileus that is not associated with cystic fibrosis (CF) is caused by different homozygous mutations in GUCY2C, leading to a dramatic reduction or fully abrogating the enzymatic activity of the encoded guanlyl cyclase 2C. GUCY2C is a transmembrane receptor whose extracellular domain is activated by either the endogenous ligands, guanylin and related peptide uroguanylin, or by an external ligand, Escherichia coli (E. coli) heat-stable enterotoxin STa. GUCY2C is expressed in the human intestine, and the encoded protein activates the CFTR protein through local generation of cGMP. Thus, GUCY2C is a likely candidate modifier of the meconium ileus phenotype in CF. Because GUCY2C heterozygous and homozygous mutant mice are resistant to E. coli STa enterotoxin-induced diarrhea, it is plausible that GUCY2C mutations in the desert-dwelling Bedouin kindred are of selective advantage. Meconium ileus (MI), intestinal obstruction by inspissated homozygosity on chromosome 12p13 (spanning 9.5 Mb meconium in the distal ileum and cecum, develops in between markers D12S366 and D12S310) that was utero and presents shortly after birth as failure to pass common to all affected individuals.
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Supp Table 1.Pdf
Upregulated genes in Hdac8 null cranial neural crest cells fold change Gene Symbol Gene Title 134.39 Stmn4 stathmin-like 4 46.05 Lhx1 LIM homeobox protein 1 31.45 Lect2 leukocyte cell-derived chemotaxin 2 31.09 Zfp108 zinc finger protein 108 27.74 0710007G10Rik RIKEN cDNA 0710007G10 gene 26.31 1700019O17Rik RIKEN cDNA 1700019O17 gene 25.72 Cyb561 Cytochrome b-561 25.35 Tsc22d1 TSC22 domain family, member 1 25.27 4921513I08Rik RIKEN cDNA 4921513I08 gene 24.58 Ofa oncofetal antigen 24.47 B230112I24Rik RIKEN cDNA B230112I24 gene 23.86 Uty ubiquitously transcribed tetratricopeptide repeat gene, Y chromosome 22.84 D8Ertd268e DNA segment, Chr 8, ERATO Doi 268, expressed 19.78 Dag1 Dystroglycan 1 19.74 Pkn1 protein kinase N1 18.64 Cts8 cathepsin 8 18.23 1500012D20Rik RIKEN cDNA 1500012D20 gene 18.09 Slc43a2 solute carrier family 43, member 2 17.17 Pcm1 Pericentriolar material 1 17.17 Prg2 proteoglycan 2, bone marrow 17.11 LOC671579 hypothetical protein LOC671579 17.11 Slco1a5 solute carrier organic anion transporter family, member 1a5 17.02 Fbxl7 F-box and leucine-rich repeat protein 7 17.02 Kcns2 K+ voltage-gated channel, subfamily S, 2 16.93 AW493845 Expressed sequence AW493845 16.12 1600014K23Rik RIKEN cDNA 1600014K23 gene 15.71 Cst8 cystatin 8 (cystatin-related epididymal spermatogenic) 15.68 4922502D21Rik RIKEN cDNA 4922502D21 gene 15.32 2810011L19Rik RIKEN cDNA 2810011L19 gene 15.08 Btbd9 BTB (POZ) domain containing 9 14.77 Hoxa11os homeo box A11, opposite strand transcript 14.74 Obp1a odorant binding protein Ia 14.72 ORF28 open reading
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(12) Patent Application Publication (10) Pub. No.: US 2003/0082511 A1 Brown Et Al
US 20030082511A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2003/0082511 A1 Brown et al. (43) Pub. Date: May 1, 2003 (54) IDENTIFICATION OF MODULATORY Publication Classification MOLECULES USING INDUCIBLE PROMOTERS (51) Int. Cl." ............................... C12O 1/00; C12O 1/68 (52) U.S. Cl. ..................................................... 435/4; 435/6 (76) Inventors: Steven J. Brown, San Diego, CA (US); Damien J. Dunnington, San Diego, CA (US); Imran Clark, San Diego, CA (57) ABSTRACT (US) Correspondence Address: Methods for identifying an ion channel modulator, a target David B. Waller & Associates membrane receptor modulator molecule, and other modula 5677 Oberlin Drive tory molecules are disclosed, as well as cells and vectors for Suit 214 use in those methods. A polynucleotide encoding target is San Diego, CA 92121 (US) provided in a cell under control of an inducible promoter, and candidate modulatory molecules are contacted with the (21) Appl. No.: 09/965,201 cell after induction of the promoter to ascertain whether a change in a measurable physiological parameter occurs as a (22) Filed: Sep. 25, 2001 result of the candidate modulatory molecule. Patent Application Publication May 1, 2003 Sheet 1 of 8 US 2003/0082511 A1 KCNC1 cDNA F.G. 1 Patent Application Publication May 1, 2003 Sheet 2 of 8 US 2003/0082511 A1 49 - -9 G C EH H EH N t R M h so as se W M M MP N FIG.2 Patent Application Publication May 1, 2003 Sheet 3 of 8 US 2003/0082511 A1 FG. 3 Patent Application Publication May 1, 2003 Sheet 4 of 8 US 2003/0082511 A1 KCNC1 ITREXCHO KC 150 mM KC 2000000 so 100 mM induced Uninduced Steady state O 100 200 300 400 500 600 700 Time (seconds) FIG.
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Supplementary Table 2
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Strand Breaks for P53 Exon 6 and 8 Among Different Time Course of Folate Depletion Or Repletion in the Rectosigmoid Mucosa
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Guanylyl Cyclase C Hormone Axis at the Intersection of Obesity and Colorectal Cancer
1521-0111/90/3/199–204$25.00 http://dx.doi.org/10.1124/mol.115.103192 MOLECULAR PHARMACOLOGY Mol Pharmacol 90:199–204, September 2016 Copyright ª 2016 by The American Society for Pharmacology and Experimental Therapeutics MINIREVIEW Guanylyl Cyclase C Hormone Axis at the Intersection of Obesity and Colorectal Cancer Erik S. Blomain, Dante J. Merlino, Amanda M. Pattison, Adam E. Snook, and Scott A. Waldman Downloaded from Department of Pharmacology and Experimental Therapeutics, Thomas Jefferson University, Philadelphia, Pennsylvania Received December 29, 2015; accepted May 25, 2016 ABSTRACT Obesity has emerged as a principal cause of mortality worldwide, in mice and humans. Hormone loss reflects reversible calorie- molpharm.aspetjournals.org reflecting comorbidities including cancer risk, particularly in induced endoplasmic reticulum stress and the associated colorectum. Although this relationship is established epidemio- unfolded protein response, rather than the endocrine, adipo- logically, molecular mechanisms linking colorectal cancer and kine, or inflammatory milieu of obesity. Loss of intestinal obesity continue to be refined. Guanylyl cyclase C (GUCY2C), uroguanylin secretion silences the hypothalamic GUCY2C a membrane-bound guanylyl cyclase expressed in intestinal endocrine axis, creating a feed-forward loop contributing to epithelial cells, binds the paracrine hormones guanylin and hyperphagia in obesity. Importantly, calorie-induced guanylin uroguanylin, inducing cGMP signaling in colorectum and small loss silences the GUCY2C-cGMP paracrine axis underlying intestine, respectively. Guanylin is the most commonly lost obesity-induced epithelial dysfunction and colorectal tumor- gene product in sporadic colorectal cancer, and its universal igenesis. Indeed, genetically enforced guanylin replacement loss early in transformation silences GUCY2C, a tumor sup- eliminated diet-induced intestinal tumorigenesis in mice.
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Dema and Faust Et Al., Suppl. Material 2020.02.03
Supplementary Materials Cyclin-dependent kinase 18 controls trafficking of aquaporin-2 and its abundance through ubiquitin ligase STUB1, which functions as an AKAP Dema Alessandro1,2¶, Dörte Faust1¶, Katina Lazarow3, Marc Wippich3, Martin Neuenschwander3, Kerstin Zühlke1, Andrea Geelhaar1, Tamara Pallien1, Eileen Hallscheidt1, Jenny Eichhorst3, Burkhard Wiesner3, Hana Černecká1, Oliver Popp1, Philipp Mertins1, Gunnar Dittmar1, Jens Peter von Kries3, Enno Klussmann1,4* ¶These authors contributed equally to this work 1Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Robert- Rössle-Strasse 10, 13125 Berlin, Germany 2current address: University of California, San Francisco, 513 Parnassus Avenue, CA 94122 USA 3Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Rössle-Strasse 10, 13125 Berlin, Germany 4DZHK (German Centre for Cardiovascular Research), Partner Site Berlin, Oudenarder Strasse 16, 13347 Berlin, Germany *Corresponding author Enno Klussmann Max Delbrück Center for Molecular Medicine Berlin in the Helmholtz Association (MDC) Robert-Rössle-Str. 10, 13125 Berlin Germany Tel. +49-30-9406 2596 FAX +49-30-9406 2593 E-mail: [email protected] 1 Content 1. CELL-BASED SCREENING BY AUTOMATED IMMUNOFLUORESCENCE MICROSCOPY 3 1.1 Screening plates 3 1.2 Image analysis using CellProfiler 17 1.4 Identification of siRNA affecting cell viability 18 1.7 Hits 18 2. SUPPLEMENTARY TABLE S4, FIGURES S2-S4 20 2 1. Cell-based screening by automated immunofluorescence microscopy 1.1 Screening plates Table S1. Genes targeted with the Mouse Protein Kinases siRNA sub-library. Genes are sorted by plate and well. Accessions refer to National Center for Biotechnology Information (NCBI, BLA) entries. The siRNAs were arranged on three 384-well microtitre platres.
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Gene Symbol Accession Alias/Prev Symbol Official Full Name AAK1 NM 014911.2 KIAA1048, Dkfzp686k16132 AP2 Associated Kinase 1
Gene Symbol Accession Alias/Prev Symbol Official Full Name AAK1 NM_014911.2 KIAA1048, DKFZp686K16132 AP2 associated kinase 1 (AAK1) AATK NM_001080395.2 AATYK, AATYK1, KIAA0641, LMR1, LMTK1, p35BP apoptosis-associated tyrosine kinase (AATK) ABL1 NM_007313.2 ABL, JTK7, c-ABL, p150 v-abl Abelson murine leukemia viral oncogene homolog 1 (ABL1) ABL2 NM_007314.3 ABLL, ARG v-abl Abelson murine leukemia viral oncogene homolog 2 (arg, Abelson-related gene) (ABL2) ACVR1 NM_001105.2 ACVRLK2, SKR1, ALK2, ACVR1A activin A receptor ACVR1B NM_004302.3 ACVRLK4, ALK4, SKR2, ActRIB activin A receptor, type IB (ACVR1B) ACVR1C NM_145259.2 ACVRLK7, ALK7 activin A receptor, type IC (ACVR1C) ACVR2A NM_001616.3 ACVR2, ACTRII activin A receptor ACVR2B NM_001106.2 ActR-IIB activin A receptor ACVRL1 NM_000020.1 ACVRLK1, ORW2, HHT2, ALK1, HHT activin A receptor type II-like 1 (ACVRL1) ADCK1 NM_020421.2 FLJ39600 aarF domain containing kinase 1 (ADCK1) ADCK2 NM_052853.3 MGC20727 aarF domain containing kinase 2 (ADCK2) ADCK3 NM_020247.3 CABC1, COQ8, SCAR9 chaperone, ABC1 activity of bc1 complex like (S. pombe) (CABC1) ADCK4 NM_024876.3 aarF domain containing kinase 4 (ADCK4) ADCK5 NM_174922.3 FLJ35454 aarF domain containing kinase 5 (ADCK5) ADRBK1 NM_001619.2 GRK2, BARK1 adrenergic, beta, receptor kinase 1 (ADRBK1) ADRBK2 NM_005160.2 GRK3, BARK2 adrenergic, beta, receptor kinase 2 (ADRBK2) AKT1 NM_001014431.1 RAC, PKB, PRKBA, AKT v-akt murine thymoma viral oncogene homolog 1 (AKT1) AKT2 NM_001626.2 v-akt murine thymoma viral oncogene homolog 2 (AKT2) AKT3 NM_181690.1
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Page 1 of 19 Diabetes Effect of guanylate cyclase-C activity on energy and glucose homeostasis Denovan P. Begg1,2, Kris A. Steinbrecher3, Joram D. Mul1, Adam P. Chambers1, Rohit Kohli3, April Haller1, Mitchell B. Cohen3, Stephen C. Woods1 and Randy J. Seeley1 1 Metabolic Diseases Institute, University of Cincinnati, Cincinnati, OH, 45237 2 School of Psychology, University of New South Wales, UNSW Sydney, NSW, Australia, 2052 3 Division of Gastroenterology, Hepatology, and Nutrition, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45220 Running title: GC-C activation and energy balance Key words: obesity, glucose tolerance, uroguanylin, guanylate cyclase-C Word count: 2143 Number of figures: 3 Corresponding Author Denovan P. Begg School of Psychology, University of New South Wales, UNSW Sydney, NSW, Australia, 2052 Ph: +61-2-9385-2441 Fax: +61-2-9385-3641 Diabetes Publish Ahead of Print, published online June 4, 2014 Diabetes Page 2 of 19 Abstract Uroguanylin is a gastrointestinal hormone primarily involved in fluid and electrolyte handling. It has recently been reported that prouroguanylin, secreted postprandially, is converted to uroguanylin in the brain and activates the receptor guanylate cyclase-C (GC- C) to reduce food intake and prevent obesity. Here, we tested CNS administration of two GC-C agonists and found no significant reduction of food intake. We also carefully phenotyped mice lacking the GC-C receptor and found them to have normal body weight, adiposity and glucose tolerance. Interestingly, uroguanylin knockout mice had a small but significant increase in body weight and adiposity that was accompanied by glucose intolerance. Our data indicate that the modest effects of uroguanylin on energy and glucose homeostasis are not centrally mediated by central GC-C receptors.
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Guanylate Cyclase C Reduces Invasion of Intestinal Epithelial Cells by Bacterial Pathogens Received: 11 October 2017 Surya Amarachintha1, Eleana Harmel-Laws1 & Kris A
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GDNF and GUCY2C Mutation Associated with Hirschsprung's Disease in a Trio Pedigree: a Case Report
GDNF and GUCY2C mutation associated with Hirschsprung's disease in a trio pedigree: a case report Senmao Chai Xiangyang No.1 People's Hospital Kai Deng Hubei University of Medicine Xiaodong Sun Xiangyang No.1 People's Hostipal Qiang Zhao Xiangyang No.1 People's Hospital Pan Fu Xiangyang No.1 People's Hospital Qianqian Xu Xiangyang No.1 People's Hospital Ming Sang ( [email protected] ) Hubei University of Medicine https://orcid.org/0000-0001-7862-0603 Case report Keywords: Hirschsprung's disease, Exome sequencing, GDNF, GUCY2C, Ganglion cells Posted Date: April 6th, 2020 DOI: https://doi.org/10.21203/rs.3.rs-21172/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 1/12 Abstract Background: Hirschsprung's disease (HSCR) caused by absence or dysfunction of ganglion cells in the submucosal plexus and intermuscular plexus of the intestinal wall. Studies have shown that homozygous mutations of glial cell-line derived neurotrophic factor ( GDNF ) can lead to the occurrence of HSCR. but heterozygous mutation must combine with other related abnormal genes could to cause the HSCR. The other Guanylyl cyclase C ( GUCY2C ) gene is rarely reported as a direct cause of HSCR. Our ndings Provides a new perspective for application of mutation screening in the genetic counseling. Case presentation: We report clinical and genetic ndings of a trio pedigree with Hirschsprung's disease. Whole exome sequencing and Sanger sequencing were performed for propositus and all members of trio pedigree, respectively. We also collected 74 healthy control samples and used Sanger sequencing to identify that found the mutation.
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