DOCSLIB.ORG

Anti-EIF2C1/AGO1 Code No

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Anti-EIF2C1/AGO1 Code No For Research Use Only. RN028P Page 1 of 4 Not for use in diagnostic procedures. RIP-Certified Antibody Anti-EIF2C1/AGO1 Code No. Quantity Concentration Form RN028P 200 µµµL 1 mg/mL Affinity Purified BACKGROUND: Eukaryotic translation initiation REACTIVITY: This antibody reacts with human and factor 2C, subunit 1 (EIF2C1; AGO1) is a member of the mouse EIF2C1 on Western blotting, Immunoprecipitation argonaute protein family. It contains Piwi/Argonaute/Zwille and RNP Immunoprecipitation. (PAZ) and Piwi domain. EIF2C1 and EIF2C2 form the RNA-induced silencing complex that mediates gene APPLICATIONS: silencing by RNA interference. EIF2C1 is also known as RNP Immunoprecipitation ; 15 µg/500 µL of cell extract the initiator of RNA-induced transcriptional gene silencing from 1.3 x 10 7 cells (TGS) in human cells. EIF2C1, DNA methyltransferase 3a, Western blotting ; 1:1,000 for chemiluminescence detection and histone deacetylase I form the transcriptional silencing system complex. EIF2C1 binds to RNA polymerase II and Immunoprecipitation ; 5 µg/500 µL of cell extract from mediates TGS through heterochromatin formation. In 5 x 10 6 cells humans, the gene encoding EIF2C1 is localized on Immunohistochemistry ; Not tested chromosome 1; this region is often deleted in cases of Immunocytochemistry ; Not tested Wilms’ tumors. EIF2C1 may be involved in tumor Flow cytometry ; Not tested development because its expression is upregulated in tumors that lack the Wilm’s tumor suppressor gene, namely, Detailed procedures are provided in the following WT1. PROTOCOLS . RIP-CERTIFIED ANTIBODY: INTENDED USE: Posttranscriptional regulation of gene expression is a For Research Use Only. Not for use in diagnostic procedures. ribonucleoprotein-driven process, which involves RNA binding proteins (RBPs) and non-coding RNAs that affect splicing, nuclear export, subcellular localization, mRNA REFERENCES: decay and translation. The RNP Immunoprecipitation-Chip 1) Kim, D. H., et al., Nat. Struct. Mol. Biol. 13, 793-797 (2006) (RIP-Chip), RIP-Seq and RIP-RTPCR allow the 2) Hawkins, P. G., et al., Nucleic Acids Res. 37, 2984-2995 (2009) identification of multiple RNA targets of RBPs globally 3) Martinez, J., et al., Cell 110, 563-574 (2002) and within the context of a cell extract. Antibodies specific 4) Carmell, M. A., et al., Genes Dev. 16, 2733-2742 (2002) to the RNA binding protein of interest are used to co-immunoprecipitate the RNA binding protein and the SPECIES CROSS REACTIVITY: associated subset of RNAs. The RNA content is interrogated using standard microarray or sequencing Species Human Mouse Rat Hamster technology. RIP-Certified Antibody is validated for use in 293T, HeLa Cell NIH/3T3 Rat1 Not Tested RNP Immunoprecipitation (RIP) in conjunction with the K562, Jurkat RIP-Assay Kit for microRNA distributed from MBL. Its ability to immunoprecipitate RNAs and RBPs complex was Reactivity on IP + + − confirmed by quantitative and qualitative analysis on NanoDrop, Bioanalyzer and RT-PCR or microarray. LICENSING OPPORTUNITY: The RIP-Assay uses SOURCE: This antibody was purified from rabbit serum patented technology (US patent No. 6,635,422, US patent by affinity column chromatography. The rabbit was No. 7,504,210) of Ribonomics, Inc. MBL manufactures and immunized with KLH conjugated synthetic peptide, distributes this product under license from Ribonomics, Inc. corresponding to N-terminus of human EIF2C1. Researchers may use this product for the purposes of their own research. Researchers are not allowed to use this product or RIP-Assay technology for commercial purpose FORMULATION: 200 µL volume of PBS containing without acquiring a license. For commercial use and 50% glycerol, pH 7.2. No preservative is contained. licensing opportunities, please contact us at [email protected] STORAGE: This antibody solution is stable for one year o from the date of purchase when stored at -20 C. MEDICAL & BIOLOGICAL LABORATORIES CO., LTD. URL https://ruo.mbl.co.jp/je/rip-assay/ e-mail [email protected], TEL 052-238-1904 RN028P Page 2 of 4 7) Add 500 µL of cell lysate (precleared sample of step 4), nt 1 2 o then incubate with gentle agitation for 3 hours at 4 C. 100 8) Wash the beads 4 times with mi-Wash Buffer (+) 75 (centrifuge the tube at 2,000 x g for 1 minute). 9) Add 250 µL of Master mix solution (mi-Solution I: 50 mi-Solution II = 10 µL: 240 µL). Vortex thoroughly, then spin-down. 40 10) Add 150 µL of mi-Solution III. Vortex thoroughly. 30 11) Centrifuge the tube at 2,000 x g for 2 minutes. 12) Transfer the supernatant to the fresh tube containing 2 miRNA 20 µL of mi-Solution IV. 13) Add 300 µL of ice-cold 2-propanol, vortex for 10 seconds. Place at -20 oC for 20 minutes. Centrifuge the tube at 12,000 x g for 10 minutes. Analysis of isolated small RNA (including miRNA) from Jurkat by silver staining 14) Transfer the supernatant, which contains small RNAs, to following denaturing PAGE. the tube containing 2 µL of mi-Solution IV. Isolation Lane1: Normal Rabbit IgG method for small RNAs from the supernatant is Lane2: RN028P described in the following step 15. In case of purification for large RNAs in the pellet, skip to step 16. 15) Add 500 µL of ice-cold 2-propanol, vortex for 10 PROTOCOLS: seconds, then place at -20 oC for 20 minutes. Centrifuge RNP Immunoprecipitation the tube at 12,000 x g for 10 minutes. Some buffers and reagents are included in the RIP-Assay Kit 16) Wash the pellet 2 times with 0.5 mL of ice-cold 70% for microRNA (code. RN1005). Please also refer to the ethanol and dry up the pellet for 5-15 minutes. protocol packaged in the RIP-Assay Kit for microRNA . 17) Dissolve the pellets in nuclease-free water. [Material Preparation] (Positive control for RNP Immunoprecipitation; Jurkat) 1. mi-Lysis Buffer (+) Before using the mi-Lysis Buffer, protease inhibitors, 1 2 3 4 5 RNase inhibitors, and DTT are added to the mi-Lysis kDa Buffer at the appropriate concentration. 2. mi-Wash Buffer (+) 150 Before using the mi-Wash Buffer, DTT is added to the mi-Wash Buffer at the appropriate concentration. 100 EIF2C1 Protocol (RNA isolation: Separation method) 1) Wash 1.3 x 10 7 cells 4 times with PBS and resuspend 75 them with 500 µL of ice-cold mi-Lysis Buffer (+) containing appropriate protease inhibitors, RNase inhibitors, and DTT. Vortex thoroughly, then incubate it Western blot analysis of EIF2C1 on ice for 10 minutes. expression in 293T (1), HeLa (2), K562 2) Centrifuge the tube at 12,000 x g for 5 minutes at 4 oC (3), Jurkat (4) and NIH/3T3 (5) using and transfer the supernatant to another tube. RN028P. 3) Add 30 µL of 50% protein A agarose beads slurry resuspended in mi-Lysis Buffer (+) into the supernatant. Incubate it at 4 oC with rotating for 1 hour. SDS-PAGE & Western Blotting 7 4) Centrifuge the tube at 2,000 x g for 1 minute at 4 oC and 1) Wash cells (approximately 1 x 10 cells) 3 times with transfer the supernatant to another fresh tube (precleared PBS and suspend them in 1 mL of Laemmli’s sample sample). buffer. 5) Mix 30 µL of 50% protein A agarose beads slurry 2) Boil the samples for 2 minutes and centrifuge. Load 10 resuspended in nuclease-free PBS with Normal Rabbit µL of sample per lane on a 1-mm-thick IgG (RIP-Assay Kit for microRNA ) or SDS-polyacrylamide gel and carry out electrophoresis. anti-EIF2C1/AGO1 polyclonal antibody at the 3) Blot the protein to a polyvinylidene difluoride (PVDF) 2 concentration suggested in the APPLICATIONS, and membrane at 1 mA/cm for 1 hour in a semi-dry transfer then add 1 mL of mi-Wash Buffer (+) into each tube. system (Transfer Buffer: 25 mM Tris, 190 mM glycine, Incubate with gently agitation for 1 hour at 4 oC. 20% MeOH). See the manufacturer's manual for precise 6) Wash the beads once with ice-cold mi-Lysis Buffer (+) transfer procedure. (centrifuge the tube at 2,000 x g for 1 minute). Carefully 4) To reduce nonspecific binding, soak the membrane in discard the supernatant using a pipettor without 10% skimmed milk (in PBS, pH 7.2) for 1 hour at room o disturbing the beads. temperature, or overnight at 4 C. RN028P Page 3 of 4 5) Incubate the membrane with primary antibody diluted 6) Wash the beads once with ice-cold Lysis Buffer with PBS, pH 7.2 containing 1% skimmed milk as (centrifuge the tube at 2,000 x g for 1 minute). Carefully suggested in the APPLICATIONS for 1 hour at room discard the supernatant using a pipettor without temperature. (The concentration of antibody will depend disturbing the beads. on the conditions.) 7) Add 500 µL of cell lysate (precleared sample of step 4), 6) Wash the membrane with PBS-T [0.05% Tween-20 in then incubate with gentle agitation for 3 hours at 4 oC PBS] (5 minutes x 3 times). 8) Wash the beads 4 times with Lysis Buffer (centrifuge the 7) Incubate the membrane with the 1:10,000 tube at 2,000 x g for 1 minute). HRP-conjugated anti-rabbit IgG (MBL; code no. 458) 9) Resuspend the beads in 20 µL of Laemmli’s sample diluted with 1% skimmed milk (in PBS, pH 7.2) for 1 buffer, boil for 3-5 minutes, and centrifuge for 5 minutes. hour at room temperature. Use 20 µL/lane for the SDS-PAGE analysis. 8) Wash the membrane with PBS-T (5 minutes x 3 times). (See SDS-PAGE & Western blotting .) 9) Wipe excess buffer off the membrane, and incubate membrane with an appropriate chemiluminescence (Positive control for Immunoprecipitation; 293T) reagent for 1 minute.
Load more

Recommended publications
  • Characterization of a 7.6-Mb Germline Deletion Encompassing the NF1 Locus and About a Hundred Genes in an NF1 Contiguous Gene Syndrome Patient
    European Journal of Human Genetics (2008) 16, 1459–1466 & 2008 Macmillan Publishers Limited All rights reserved 1018-4813/08 $32.00 www.nature.com/ejhg ARTICLE Characterization of a 7.6-Mb germline deletion encompassing the NF1 locus and about a hundred genes in an NF1 contiguous gene syndrome patient Eric Pasmant*,1,2, Aure´lie de Saint-Trivier2, Ingrid Laurendeau1, Anne Dieux-Coeslier3, Be´atrice Parfait1,2, Michel Vidaud1,2, Dominique Vidaud1,2 and Ivan Bie`che1,2 1UMR745 INSERM, Universite´ Paris Descartes, Faculte´ des Sciences Pharmaceutiques et Biologiques, Paris, France; 2Service de Biochimie et de Ge´ne´tique Mole´culaire, Hoˆpital Beaujon AP-HP, Clichy, France; 3Service de Ge´ne´tique Clinique, Hoˆpital Jeanne de Flandre, Lille, France We describe a large germline deletion removing the NF1 locus, identified by heterozygosity mapping based on microsatellite markers, in an 8-year-old French girl with a particularly severe NF1 contiguous gene syndrome. We used gene-dose mapping with sequence-tagged site real-time PCR to locate the deletion end points, which were precisely characterized by means of long-range PCR and nucleotide sequencing. The deletion is located on chromosome arm 17q and is exactly 7 586 986 bp long. It encompasses the entire NF1 locus and about 100 other genes, including numerous chemokine genes, an attractive in silico-selected cerebrally expressed candidate gene (designated NUFIP2, for nuclear fragile X mental retardation protein interacting protein 2; NM_020772) and four microRNA genes. Interestingly, the centromeric breakpoint is located in intron 4 of the PIPOX gene (pipecolic acid oxidase; NM_016518) and the telomeric breakpoint in intron 5 of the GGNBP2 gene (gametogenetin binding protein 2; NM_024835) coding a transcription factor.
    [Show full text]
  • Download Article (PDF)
    Article in press - uncorrected proof BioMol Concepts, Vol. 2 (2011), pp. 343–352 • Copyright ᮊ by Walter de Gruyter • Berlin • Boston. DOI 10.1515/BMC.2011.033 Review Fragile X family members have important and non-overlapping functions Claudia Winograd2 and Stephanie Ceman1,2,* member, FMR1, was isolated by positional cloning of the X 1 Department of Cell and Developmental Biology, chromosomal region containing the inducible fragile site in University of Illinois, 601 S. Goodwin Avenue, individuals with fragile X syndrome (1). Cloning of the gene Urbana–Champaign, IL 61801, USA revealed the molecular defect to be a trinucleotide (CGG) 2 Neuroscience Program and College of Medicine, repeat expansion in exon 1 (2). Normally, individuals have University of Illinois, 601 S. Goodwin Avenue, less than 45 repeats with an average around 30 repeats; how- Urbana–Champaign, IL 61801, USA ever, expansion to greater than 200 repeats leads to aberrant methylation of the cytosines, leading to recruitment of * Corresponding author histone deacetylases with consequent transcriptional silenc- e-mail: [email protected] ing of the FMR1 locus (3). Thus, individuals with fragile X syndrome do not express transcript from the FMR1 locus. Abstract To identify the Xenopus laevis ortholog of FMR1 for further use in developmental studies, the human FMR1 gene was The fragile X family of genes encodes a small family of used to screen a cDNA library prepared from Xenopus laevis RNA binding proteins including FMRP, FXR1P and FXR2P ovary. In addition to identifying the Xenopus laevis ortholog that were identified in the 1990s. All three members are of FMR1, the first autosomal paralog FXR1 was discovered encoded by 17 exons and show alternative splicing at the 39 because of its sequence similarity to FMR1 (4).
    [Show full text]
  • Anti-EIF2C1 / AGO1 Antibody (ARG63916)
    Product datasheet [email protected] ARG63916 Package: 100 μg anti-EIF2C1 / AGO1 antibody Store at: -20°C Summary Product Description Goat Polyclonal antibody recognizes EIF2C1 / AGO1 Tested Reactivity Hu Predict Reactivity Ms, Rat, Dog Tested Application IHC-P, WB Specificity This product is not expected to cross-react with EIF2C2, EIF2C3 and EIF2C4. Host Goat Clonality Polyclonal Isotype IgG Target Name EIF2C1 / AGO1 Antigen Species Human Immunogen C-KNASYNLDPYIQEF Conjugation Un-conjugated Alternate Names GERP95; Q99; eIF2C 1; EIF2C; EIF2C1; Argonaute RISC catalytic component 1; Argonaute1; Protein argonaute-1; hAgo1; Putative RNA-binding protein Q99; eIF-2C 1; Eukaryotic translation initiation factor 2C 1 Application Instructions Application table Application Dilution IHC-P 2.5 µg/ml WB 0.3 - 1 µg/ml Application Note WB: Recommend incubate at RT for 1h. IHC-P: Antigen Retrieval: Steam tissue section in Citrate buffer (pH 6.0). * The dilutions indicate recommended starting dilutions and the optimal dilutions or concentrations should be determined by the scientist. Calculated Mw 97 kDa Properties Form Liquid Purification Purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide. Buffer Tris saline (pH 7.3), 0.02% Sodium azide and 0.5% BSA Preservative 0.02% Sodium azide Stabilizer 0.5% BSA www.arigobio.com 1/3 Concentration 0.5 mg/ml Storage instruction For continuous use, store undiluted antibody at 2-8°C for up to a week. For long-term storage, aliquot and store at -20°C or below. Storage in frost free freezers is not recommended. Avoid repeated freeze/thaw cycles.
    [Show full text]
  • Gene Expression During Normal and FSHD Myogenesis Tsumagari Et Al
    Gene expression during normal and FSHD myogenesis Tsumagari et al. Tsumagari et al. BMC Medical Genomics 2011, 4:67 http://www.biomedcentral.com/1755-8794/4/67 (27 September 2011) Tsumagari et al. BMC Medical Genomics 2011, 4:67 http://www.biomedcentral.com/1755-8794/4/67 RESEARCHARTICLE Open Access Gene expression during normal and FSHD myogenesis Koji Tsumagari1, Shao-Chi Chang1, Michelle Lacey2,3, Carl Baribault2,3, Sridar V Chittur4, Janet Sowden5, Rabi Tawil5, Gregory E Crawford6 and Melanie Ehrlich1,3* Abstract Background: Facioscapulohumeral muscular dystrophy (FSHD) is a dominant disease linked to contraction of an array of tandem 3.3-kb repeats (D4Z4) at 4q35. Within each repeat unit is a gene, DUX4, that can encode a protein containing two homeodomains. A DUX4 transcript derived from the last repeat unit in a contracted array is associated with pathogenesis but it is unclear how. Methods: Using exon-based microarrays, the expression profiles of myogenic precursor cells were determined. Both undifferentiated myoblasts and myoblasts differentiated to myotubes derived from FSHD patients and controls were studied after immunocytochemical verification of the quality of the cultures. To further our understanding of FSHD and normal myogenesis, the expression profiles obtained were compared to those of 19 non-muscle cell types analyzed by identical methods. Results: Many of the ~17,000 examined genes were differentially expressed (> 2-fold, p < 0.01) in control myoblasts or myotubes vs. non-muscle cells (2185 and 3006, respectively) or in FSHD vs. control myoblasts or myotubes (295 and 797, respectively). Surprisingly, despite the morphologically normal differentiation of FSHD myoblasts to myotubes, most of the disease-related dysregulation was seen as dampening of normal myogenesis- specific expression changes, including in genes for muscle structure, mitochondrial function, stress responses, and signal transduction.
    [Show full text]
  • Small Rnas in Early Mammalian Development: from Gametes to Gastrulation Nayoung Suh and Robert Blelloch*
    REVIEW 1653 Development 138, 1653-1661 (2011) doi:10.1242/dev.056234 © 2011. Published by The Company of Biologists Ltd Small RNAs in early mammalian development: from gametes to gastrulation Nayoung Suh and Robert Blelloch* Summary Classes and biogenesis of mammalian small RNAs Small non-coding RNAs, including microRNAs (miRNAs), Three major classes of functional small non-coding RNAs have endogenous small interfering RNAs (endo-siRNAs) and Piwi- been found in mammals: microRNAs (miRNAs), endogenous interacting RNAs (piRNAs), play essential roles in mammalian small interfering RNAs (endo-siRNAs) and Piwi-interacting RNAs development. The function and timing of expression of these (piRNAs) (Babiarz and Blelloch, 2009; Kim et al., 2009; Thomson three classes of small RNAs differ greatly. piRNAs are expressed and Lin, 2009). These classes differ in their biogenesis, i.e. their and play a crucial role during male gametogenesis, whereas maturation from transcribed forms to the active form of the RNA endo-siRNAs are essential for oocyte meiosis. By contrast, (Fig. 1). miRNAs are ubiquitously expressed in somatic tissues and miRNAs can be divided into two subclasses: canonical and non- function throughout post-implantation development. canonical miRNAs. Canonical miRNAs are initially transcribed as Surprisingly, however, miRNAs are non-essential during pre- long RNAs that contain hairpins (Fig. 1A). The 60-75 nt hairpins implantation embryonic development and their function is are recognized by the RNA-binding protein Dgcr8 (DiGeorge suppressed during oocyte meiosis. Here, we review the roles of syndrome critical region 8), which directs the RNase III enzyme small non-coding RNAs during the early stages of mammalian Drosha to cleave the base of the hairpin (Denli et al., 2004; development, from gamete maturation through to Gregory et al., 2004; Han et al., 2004; Han et al., 2006; Landthaler gastrulation.
    [Show full text]
  • A Fragile X Mental Retardation-Like Gene in a Cnidarian
    Gene 343 (2004) 231–238 www.elsevier.com/locate/gene A fragile X mental retardation-like gene in a cnidarian Jasenka Guduric-Fuchsa, Frank Mfhrlena, Marcus Frohmeb, Uri Franka,* aInstitute of Zoology, University of Heidelberg, INF 230, Heidelberg 69120, Germany bDepartment of Functional Genome Analysis, German Cancer Research Center, INF 580, 69120 Heidelberg, Germany Received 6 August 2004; received in revised form 9 September 2004; accepted 5 October 2004 Available online 10 November 2004 Received by D.A. Tagle Abstract The fragile X mental retardation syndrome in humans is caused by a mutational loss of function of the fragile X mental retardation gene 1 (FMR1). FMR1 is an RNA-binding protein, involved in the development and function of the nervous system. Despite of its medical significance, the evolutionary origin of FMR1 has been unclear. Here, we report the molecular characterization of HyFMR1, an FMR1 orthologue, from the cnidarian hydroid Hydractinia echinata. Cnidarians are the most basal metazoans possessing neurons. HyFMR1is expressed throughout the life cycle of Hydractinia. Its expression pattern correlates to the position of neurons and their precursor stem cells in the animal. Our data indicate that the origin of the fraxile X related (FXR) protein family dates back at least to the common ancestor of cnidarians and bilaterians. The lack of FXR proteins in other invertebrates may have been due to gene loss in particular lineages. D 2004 Elsevier B.V. All rights reserved. Keywords: FMR1; FMRP; FXR; Hydractinia; Evolution; Hydrozoa 1. Introduction three types of RNA binding motifs: a ribonucleoprotein K homology domain (KH domain; FMR1 has two such The fragile X syndrome is the most common form of domains), an arginine and glycine rich domain (RGG box) inherited mental retardation in humans.
    [Show full text]
  • Gene Ontology Functional Annotations and Pleiotropy
    Network based analysis of genetic disease associations Sarah Gilman Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy under the Executive Committee of the Graduate School of Arts and Sciences COLUMBIA UNIVERSITY 2014 © 2013 Sarah Gilman All Rights Reserved ABSTRACT Network based analysis of genetic disease associations Sarah Gilman Despite extensive efforts and many promising early findings, genome-wide association studies have explained only a small fraction of the genetic factors contributing to common human diseases. There are many theories about where this “missing heritability” might lie, but increasingly the prevailing view is that common variants, the target of GWAS, are not solely responsible for susceptibility to common diseases and a substantial portion of human disease risk will be found among rare variants. Relatively new, such variants have not been subject to purifying selection, and therefore may be particularly pertinent for neuropsychiatric disorders and other diseases with greatly reduced fecundity. Recently, several researchers have made great progress towards uncovering the genetics behind autism and schizophrenia. By sequencing families, they have found hundreds of de novo variants occurring only in affected individuals, both large structural copy number variants and single nucleotide variants. Despite studying large cohorts there has been little recurrence among the genes implicated suggesting that many hundreds of genes may underlie these complex phenotypes. The question
    [Show full text]
  • WO 2012/170711 Al 13 December 2012 (13.12.2012) P O P C T
    (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2012/170711 Al 13 December 2012 (13.12.2012) P O P C T (51) International Patent Classification: (72) Inventors; and G01N33/5 74 (2006.01) (75) Inventors/Applicants (for US only): PAWLOWSKI, Traci [US/US]; 2014 N Milkweed Loop, Phoenix, AZ (21) International Application Number: 85037 (US). YEATTS, Kimberly [US/US]; 109 E. Pierce PCT/US2012/041387 Street, Tempe, AZ 85281 (US). AKHAVAN, Ray (22) International Filing Date: [US/US]; 5804 Malvern Hill Ct, Haymarket, VA 201 69 7 June 2012 (07.06.2012) (US). (25) Filing Language: English (74) Agent: AKHAVAN, Ramin; Caris Science, Inc., 6655 N. MacArthur Blvd., Irving, TX 75039 (US). (26) Publication Language: English (81) Designated States (unless otherwise indicated, for every (30) Priority Data: kind of national protection available): AE, AG, AL, AM, 61/494,196 7 June 201 1 (07.06.201 1) AO, AT, AU, AZ, BA, BB, BG, BH, BR, BW, BY, BZ, 61/494,355 7 June 201 1 (07.06.201 1) CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, DO, 61/507,989 14 July 201 1 (14.07.201 1) DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, (71) Applicant (for all designated States except US): CARIS HR, HU, ID, IL, IN, IS, JP, KE, KG, KM, KN, KP, KR, LIFE SCIENCES LUXEMBOURG HOLDINGS, KZ, LA, LC, LK, LR, LS, LT, LU, LY, MA, MD, ME, S.A.R.L [LU/LU]; Rue De Maraichers, L2124 Luxem MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, bourg, Grand-Duche De Luxembourg (LU).
    [Show full text]
  • WO 2012/174282 A2 20 December 2012 (20.12.2012) P O P C T
    (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2012/174282 A2 20 December 2012 (20.12.2012) P O P C T (51) International Patent Classification: David [US/US]; 13539 N . 95th Way, Scottsdale, AZ C12Q 1/68 (2006.01) 85260 (US). (21) International Application Number: (74) Agent: AKHAVAN, Ramin; Caris Science, Inc., 6655 N . PCT/US20 12/0425 19 Macarthur Blvd., Irving, TX 75039 (US). (22) International Filing Date: (81) Designated States (unless otherwise indicated, for every 14 June 2012 (14.06.2012) kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BR, BW, BY, BZ, English (25) Filing Language: CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, DO, Publication Language: English DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IS, JP, KE, KG, KM, KN, KP, KR, (30) Priority Data: KZ, LA, LC, LK, LR, LS, LT, LU, LY, MA, MD, ME, 61/497,895 16 June 201 1 (16.06.201 1) US MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, 61/499,138 20 June 201 1 (20.06.201 1) US OM, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SC, SD, 61/501,680 27 June 201 1 (27.06.201 1) u s SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, 61/506,019 8 July 201 1(08.07.201 1) u s TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW.
    [Show full text]
  • Changes in Mirna Expression in a Model of Microcephaly Shan Parikh University of Connecticut - Storrs, [email protected]
    University of Connecticut OpenCommons@UConn Honors Scholar Theses Honors Scholar Program Spring 5-9-2010 Changes in miRNA Expression in a Model of Microcephaly Shan Parikh University of Connecticut - Storrs, [email protected] Follow this and additional works at: https://opencommons.uconn.edu/srhonors_theses Part of the Cellular and Molecular Physiology Commons, and the Other Physiology Commons Recommended Citation Parikh, Shan, "Changes in miRNA Expression in a Model of Microcephaly" (2010). Honors Scholar Theses. 140. https://opencommons.uconn.edu/srhonors_theses/140 Changes in miRNA expression in a model of Microcephaly Shan Parikh Physiology and Neurobiology Abstract miRNAs function to regulate gene expression through post-transcriptional mechanisms to potentially regulate multiple aspects of physiology and development. Whole transcriptome analysis has been conducted on the citron kinase mutant rat, a mutant that shows decreases in brain growth and development. The resulting differences in RNA between mutant and wild-type controls can be used to identify genetic pathways that may be regulated differentially in normal compared to abnormal neurogenesis. The goal of this thesis was to verify, with quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), changes in miRNA expression in Cit-k mutants and wild types. In addition to confirming miRNA expression changes, bio- informatics software TargetScan 5.1 was used to identify potential mRNA targets of the differentially expressed miRNAs. The miRNAs that were confirmed to change include: rno-miR- 466c, mmu-miR-493, mmu-miR-297a, hsa-miR-765, and hsa-miR-1270. The TargetScan analysis revealed 347 potential targets which have known roles in development. A subset of these potential targets include genes involved in the Wnt signaling pathway which is known to be an important regulator of stem cell development.
    [Show full text]
  • WO2011085347A2.Pdf
    (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date 14 July 2011 (14.07.2011) WO 2011/085347 A2 (51) International Patent Classification: AO, AT, AU, AZ, BA, BB, BG, BH, BR, BW, BY, BZ, C12N 15/113 (2010.01) A61K 48/00 (2006.01) CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, DO, A61K 31/7088 (2006.01) C12N 15/63 (2006.01) DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IS, JP, KE, KG, KM, KN, KP, (21) International Application Number: KR, KZ, LA, LC, LK, LR, LS, LT, LU, LY, MA, MD, PCT/US201 1/020768 ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, (22) International Filing Date: NO, NZ, OM, PE, PG, PH, PL, PT, RO, RS, RU, SC, SD, 11 January 201 1 ( 11.01 .201 1) SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (25) Filing Language: English (84) Designated States (unless otherwise indicated, for every (26) Publication Language: English kind of regional protection available): ARIPO (BW, GH, (30) Priority Data: GM, KE, LR, LS, MW, MZ, NA, SD, SL, SZ, TZ, UG, 61/293,739 11 January 2010 ( 11.01 .2010) US ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, (71) Applicant (for all designated States except US): OPKO EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, ΓΓ, LT, LU, CURNA, LLC [US/US]; 440 Biscayne Boulevard, Mia LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, mi, FL 33 137 (US).
    [Show full text]
  • The Argonaute Family: Tentacles That Reach Into Rnai, Developmental Control, Stem Cell Maintenance, and Tumorigenesis
    Downloaded from genesdev.cshlp.org on September 26, 2021 - Published by Cold Spring Harbor Laboratory Press The Argonaute family: tentacles that reach into RNAi, developmental control, stem cell maintenance, and tumorigenesis Michelle A. Carmell,1,2,3 Zhenyu Xuan,1,3 Michael Q. Zhang,1 and Gregory J. Hannon1,4 1Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724, USA; 2Program in Genetics, State University of New York at Stony Brook, Stony Brook, New York 11794, USA RNA interference (RNAi) is an evolutionarily conserved The Argonaute family process through which double-stranded RNA (dsRNA) Argonaute proteins make up a highly conserved family induces the silencing of cognate genes (for review, see whose members have been implicated in RNAi and re- Bernstein et al. 2001b; Carthew 2001). Sources of dsRNA lated phenomena in several organisms. In addition to silencing triggers include experimentally introduced roles in RNAi-like mechanisms, Argonaute proteins in- dsRNAs, RNA viruses, transposons, and RNAs tran- fluence development, and at least a subset are involved scribed from complex transgene arrays (for review, see in stem cell fate determination. Argonaute proteins are Hammond et al. 2001b). Short hairpin sequences en- ∼100-kD highly basic proteins that contain two common coded in the genome also appear to enter the RNAi path- domains, namely PAZ and PIWI domains (Cerutti et al. way and function to regulate the expression of endog- 2000). The PAZ domain, consisting of 130 amino acids, enous, protein-coding genes (Grishok et al. 2001; has been identified in Argonaute proteins and in Dicer Hutvagner et al. 2001; Ketting et al.
    [Show full text]