Mouse Dcbld2 Conditional Knockout Project (CRISPR/Cas9)

Total Page:16

File Type:pdf, Size:1020Kb

Mouse Dcbld2 Conditional Knockout Project (CRISPR/Cas9) https://www.alphaknockout.com Mouse Dcbld2 Conditional Knockout Project (CRISPR/Cas9) Objective: To create a Dcbld2 conditional knockout Mouse model (C57BL/6J) by CRISPR/Cas-mediated genome engineering. Strategy summary: The Dcbld2 gene (NCBI Reference Sequence: NM_028523 ; Ensembl: ENSMUSG00000035107 ) is located on Mouse chromosome 16. 16 exons are identified, with the ATG start codon in exon 1 and the TGA stop codon in exon 16 (Transcript: ENSMUST00000046663). Exon 5~7 will be selected as conditional knockout region (cKO region). Deletion of this region should result in the loss of function of the Mouse Dcbld2 gene. To engineer the targeting vector, homologous arms and cKO region will be generated by PCR using BAC clone RP23-86O4 as template. Cas9, gRNA and targeting vector will be co-injected into fertilized eggs for cKO Mouse production. The pups will be genotyped by PCR followed by sequencing analysis. Note: Mice homozygous for a knock-out allele exhibit reduced postnatal angiogenesis and impaired recovery from femoral artery ligation with impaired blood flow and decreased capillary density. Exon 5 starts from about 26.66% of the coding region. The knockout of Exon 5~7 will result in frameshift of the gene. The size of intron 4 for 5'-loxP site insertion: 3059 bp, and the size of intron 7 for 3'-loxP site insertion: 884 bp. The size of effective cKO region: ~1888 bp. The cKO region does not have any other known gene. Page 1 of 8 https://www.alphaknockout.com Overview of the Targeting Strategy Wildtype allele 5' gRNA region gRNA region 3' 1 5 6 7 8 9 16 Targeting vector Targeted allele Constitutive KO allele (After Cre recombination) Legends Exon of mouse Dcbld2 Homology arm cKO region loxP site Page 2 of 8 https://www.alphaknockout.com Overview of the Dot Plot Window size: 10 bp Forward Reverse Complement Sequence 12 Note: The sequence of homologous arms and cKO region is aligned with itself to determine if there are tandem repeats. No significant tandem repeat is found in the dot plot matrix. So this region is suitable for PCR screening or sequencing analysis. Overview of the GC Content Distribution Window size: 300 bp Sequence 12 Summary: Full Length(8388bp) | A(26.13% 2192) | C(19.87% 1667) | T(32.61% 2735) | G(21.39% 1794) Note: The sequence of homologous arms and cKO region is analyzed to determine the GC content. No significant high GC-content region is found. So this region is suitable for PCR screening or sequencing analysis. Page 3 of 8 https://www.alphaknockout.com BLAT Search Results (up) QUERY SCORE START END QSIZE IDENTITY CHROM STRAND START END SPAN ----------------------------------------------------------------------------------------------- browser details YourSeq 3000 1 3000 3000 100.0% chr16 + 58445203 58448202 3000 browser details YourSeq 149 825 1025 3000 90.1% chr5 - 147769073 147769252 180 browser details YourSeq 147 825 991 3000 96.9% chr4 - 59291648 59291816 169 browser details YourSeq 142 825 991 3000 93.4% chr10 + 16053684 16053844 161 browser details YourSeq 139 825 991 3000 95.9% chr17 - 51968195 51968359 165 browser details YourSeq 138 825 991 3000 95.2% chr3 - 140743622 140743786 165 browser details YourSeq 137 845 991 3000 98.0% chr11 + 112734954 112735113 160 browser details YourSeq 135 825 991 3000 91.6% chr9 - 47995644 47995796 153 browser details YourSeq 135 845 991 3000 97.9% chr4 - 58322789 58323034 246 browser details YourSeq 134 825 991 3000 90.8% chr1 - 22327761 22327909 149 browser details YourSeq 133 845 991 3000 97.2% chr8 - 95515476 95515643 168 browser details YourSeq 133 845 991 3000 93.4% chr10 + 66495089 66495224 136 browser details YourSeq 132 845 991 3000 98.6% chr9 + 48217646 48217817 172 browser details YourSeq 128 825 991 3000 97.1% chr6 + 32110081 32110253 173 browser details YourSeq 126 825 991 3000 94.4% chrY + 77732162 77732378 217 browser details YourSeq 126 825 991 3000 94.4% chrY + 65416094 65416318 225 browser details YourSeq 126 825 991 3000 94.4% chrY + 54693978 54694194 217 browser details YourSeq 125 853 991 3000 97.8% chr8 - 77820822 77820975 154 browser details YourSeq 122 1827 1989 3000 90.1% chr7 - 58629345 58629508 164 browser details YourSeq 122 871 1003 3000 97.7% chr6 - 138050451 138050628 178 Note: The 3000 bp section upstream of Exon 5 is BLAT searched against the genome. No significant similarity is found. BLAT Search Results (down) QUERY SCORE START END QSIZE IDENTITY CHROM STRAND START END SPAN ----------------------------------------------------------------------------------------------- browser details YourSeq 3000 1 3000 3000 100.0% chr16 + 58450091 58453090 3000 browser details YourSeq 44 2483 2536 3000 95.9% chr6 + 99504441 99504498 58 browser details YourSeq 42 2486 2552 3000 74.0% chr15 + 36196369 36196417 49 browser details YourSeq 31 2790 2820 3000 100.0% chr18 + 86972892 86972922 31 browser details YourSeq 31 2473 2508 3000 97.2% chr10 + 113763670 113763719 50 Note: The 3000 bp section downstream of Exon 7 is BLAT searched against the genome. No significant similarity is found. Page 4 of 8 https://www.alphaknockout.com Gene and protein information: Dcbld2 discoidin, CUB and LCCL domain containing 2 [ Mus musculus (house mouse) ] Gene ID: 73379, updated on 14-Aug-2019 Gene summary Official Symbol Dcbld2 provided by MGI Official Full Name discoidin, CUB and LCCL domain containing 2 provided by MGI Primary source MGI:MGI:1920629 See related Ensembl:ENSMUSG00000035107 Gene type protein coding RefSeq status VALIDATED Organism Mus musculus Lineage Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia; Eutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae; Murinae; Mus; Mus Also known as Esdn; CLCP1; AW146002; 1700055P21Rik Expression Ubiquitous expression in bladder adult (RPKM 10.6), CNS E11.5 (RPKM 10.5) and 27 other tissues See more Orthologs human all Genomic context Location: 16; 16 C1.2 See Dcbld2 in Genome Data Viewer Exon count: 16 Annotation release Status Assembly Chr Location 108 current GRCm38.p6 (GCF_000001635.26) 16 NC_000082.6 (58408535..58469745) Build 37.2 previous assembly MGSCv37 (GCF_000001635.18) 16 NC_000082.5 (58408648..58469858) Chromosome 16 - NC_000082.6 Page 5 of 8 https://www.alphaknockout.com Transcript information: This gene has 7 transcripts Gene: Dcbld2 ENSMUSG00000035107 Description discoidin, CUB and LCCL domain containing 2 [Source:MGI Symbol;Acc:MGI:1920629] Gene Synonyms 1700055P21Rik, CLCP1, Esdn Location Chromosome 16: 58,408,443-58,469,727 forward strand. GRCm38:CM001009.2 About this gene This gene has 7 transcripts (splice variants), 208 orthologues, 35 paralogues, is a member of 1 Ensembl protein family and is associated with 12 phenotypes. Transcripts Name Transcript ID bp Protein Translation ID Biotype CCDS UniProt Flags Dcbld2-201 ENSMUST00000046663.7 6561 769aa ENSMUSP00000039915.7 Protein coding CCDS28229 Q91ZV3 TSL:1 GENCODE basic APPRIS P1 Dcbld2-202 ENSMUST00000130409.7 2396 No protein - Retained intron - - TSL:1 Dcbld2-204 ENSMUST00000135415.1 2359 No protein - Retained intron - - TSL:1 Dcbld2-203 ENSMUST00000134324.7 891 No protein - Retained intron - - TSL:3 Dcbld2-207 ENSMUST00000150817.7 1270 No protein - lncRNA - - TSL:1 Dcbld2-205 ENSMUST00000142830.7 627 No protein - lncRNA - - TSL:3 Dcbld2-206 ENSMUST00000149321.1 576 No protein - lncRNA - - TSL:3 Page 6 of 8 https://www.alphaknockout.com 81.28 kb Forward strand 58.40Mb 58.42Mb 58.44Mb 58.46Mb Genes (Comprehensive set... Dcbld2-201 >protein coding Dcbld2-205 >lncRNA Dcbld2-206 >lncRNA Dcbld2-203 >retained intron Dcbld2-207 >lncRNA Dcbld2-202 >retained intron Dcbld2-204 >retained intron Contigs CT027564.7 > Genes < 4930461C15Rik-201lncRNA < St3gal6-207protein coding (Comprehensive set... < St3gal6-202protein coding < St3gal6-201protein coding < St3gal6-212lncRNA Regulatory Build 58.40Mb 58.42Mb 58.44Mb 58.46Mb Reverse strand 81.28 kb Regulation Legend CTCF Enhancer Open Chromatin Promoter Promoter Flank Gene Legend Protein Coding Ensembl protein coding merged Ensembl/Havana Non-Protein Coding RNA gene processed transcript Page 7 of 8 https://www.alphaknockout.com Transcript: ENSMUST00000046663 61.28 kb Forward strand Dcbld2-201 >protein coding ENSMUSP00000039... Transmembrane heli... MobiDB lite Low complexity (Seg) Superfamily Galactose-binding-like domain superfamily LCCL domain superfamily Spermadhesin, CUB domain superfamily SMART CUB domain LCCL domain Coagulation factor 5/8 C-terminal domain Pfam CUB domain LCCL domain Coagulation factor 5/8 C-terminal domain PROSITE profiles CUB domain LCCL domain Coagulation factor 5/8 C-terminal domain PROSITE patterns Coagulation factor 5/8 C-terminal domain PANTHER PTHR46806:SF3 PTHR46806 Gene3D Spermadhesin, CUB domain superfamily Galactose-binding-like domain superfamily LCCL domain superfamily CDD CUB domain Coagulation factor 5/8 C-terminal domain All sequence SNPs/i... Sequence variants (dbSNP and all other sources) Variant Legend inframe insertion missense variant splice region variant synonymous variant Scale bar 0 80 160 240 320 400 480 560 640 769 We wish to acknowledge the following valuable scientific information resources: Ensembl, MGI, NCBI, UCSC. Page 8 of 8.
Recommended publications
  • EGFR Phosphorylation of DCBLD2 Recruits TRAF6 and Stimulates AKT-Promoted Tumorigenesis
    The Journal of Clinical Investigation RESEARCH ARTICLE EGFR phosphorylation of DCBLD2 recruits TRAF6 and stimulates AKT-promoted tumorigenesis Haizhong Feng,1,2 Giselle Y. Lopez,3 Chung Kwon Kim,2 Angel Alvarez,2 Christopher G. Duncan,3 Ryo Nishikawa,4 Motoo Nagane,5 An-Jey A. Su,6 Philip E. Auron,6 Matthew L. Hedberg,7 Lin Wang,7 Jeffery J. Raizer,2 John A. Kessler,2 Andrew T. Parsa,8 Wei-Qiang Gao,1 Sung-Hak Kim,9 Mutsuko Minata,9 Ichiro Nakano,9 Jennifer R. Grandis,7 Roger E. McLendon,3 Darell D. Bigner,3 Hui-Kuan Lin,10 Frank B. Furnari,11 Webster K. Cavenee,11 Bo Hu,2 Hai Yan,3 and Shi-Yuan Cheng1,2 1State Key Laboratory of Oncogenes and Related Genes, Renji-Med X Clinical Stem Cell Research Center, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China. 2Department of Neurology and Northwestern Brain Tumor Institute, Center for Genetic Medicine, Robert H. Lurie Comprehensive Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA. 3Pediatric Brain Tumor Foundation Institute at Duke, The Preston Robert Tisch Brain Tumor Center, and Department of Pathology, Duke University Medical Center, Durham, North Carolina, USA. 4Department of Neuro-Oncology/Neurosurgery, International Medical Center, Saitama Medical University, Saitama, Japan. 5Department of Neurosurgery, Kyorin University, Tokyo, Japan. 6Department of Biological Sciences, Duquesne University, Pittsburgh, Pennsylvania, USA. 7Departments of Otolaryngology and Pharmacology and Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA. 8Department of Neurological Surgery and Northwestern Brain Tumor Institute, Robert H. Lurie Comprehensive Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.
    [Show full text]
  • Downloaded the “Top Edge” Version
    bioRxiv preprint doi: https://doi.org/10.1101/855338; this version posted December 6, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. 1 Drosophila models of pathogenic copy-number variant genes show global and 2 non-neuronal defects during development 3 Short title: Non-neuronal defects of fly homologs of CNV genes 4 Tanzeen Yusuff1,4, Matthew Jensen1,4, Sneha Yennawar1,4, Lucilla Pizzo1, Siddharth 5 Karthikeyan1, Dagny J. Gould1, Avik Sarker1, Yurika Matsui1,2, Janani Iyer1, Zhi-Chun Lai1,2, 6 and Santhosh Girirajan1,3* 7 8 1. Department of Biochemistry and Molecular Biology, Pennsylvania State University, 9 University Park, PA 16802 10 2. Department of Biology, Pennsylvania State University, University Park, PA 16802 11 3. Department of Anthropology, Pennsylvania State University, University Park, PA 16802 12 4 contributed equally to work 13 14 *Correspondence: 15 Santhosh Girirajan, MBBS, PhD 16 205A Life Sciences Building 17 Pennsylvania State University 18 University Park, PA 16802 19 E-mail: [email protected] 20 Phone: 814-865-0674 21 1 bioRxiv preprint doi: https://doi.org/10.1101/855338; this version posted December 6, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. 22 ABSTRACT 23 While rare pathogenic copy-number variants (CNVs) are associated with both neuronal and non- 24 neuronal phenotypes, functional studies evaluating these regions have focused on the molecular 25 basis of neuronal defects.
    [Show full text]
  • EGFR Phosphorylation of DCBLD2 Recruits TRAF6 and Stimulates AKT-Promoted Tumorigenesis
    EGFR phosphorylation of DCBLD2 recruits TRAF6 and stimulates AKT-promoted tumorigenesis Haizhong Feng, … , Hai Yan, Shi-Yuan Cheng J Clin Invest. 2014;124(9):3741-3756. https://doi.org/10.1172/JCI73093. Research Article Oncology Aberrant activation of EGFR in human cancers promotes tumorigenesis through stimulation of AKT signaling. Here, we determined that the discoidina neuropilin-like membrane protein DCBLD2 is upregulated in clinical specimens of glioblastomas and head and neck cancers (HNCs) and is required for EGFR-stimulated tumorigenesis. In multiple cancer cell lines, EGFR activated phosphorylation of tyrosine 750 (Y750) of DCBLD2, which is located within a recently identified binding motif for TNF receptor-associated factor 6 (TRAF6). Consequently, phosphorylation of DCBLD2 Y750 recruited TRAF6, leading to increased TRAF6 E3 ubiquitin ligase activity and subsequent activation of AKT, thereby enhancing EGFR-driven tumorigenesis. Moreover, evaluation of patient samples of gliomas and HNCs revealed an association among EGFR activation, DCBLD2 phosphorylation, and poor prognoses. Together, our findings uncover a pathway in which DCBLD2 functions as a signal relay for oncogenic EGFR signaling to promote tumorigenesis and suggest DCBLD2 and TRAF6 as potential therapeutic targets for human cancers that are associated with EGFR activation. Find the latest version: https://jci.me/73093/pdf The Journal of Clinical Investigation RESEARCH ARTICLE EGFR phosphorylation of DCBLD2 recruits TRAF6 and stimulates AKT-promoted tumorigenesis Haizhong Feng,1,2 Giselle Y. Lopez,3 Chung Kwon Kim,2 Angel Alvarez,2 Christopher G. Duncan,3 Ryo Nishikawa,4 Motoo Nagane,5 An-Jey A. Su,6 Philip E. Auron,6 Matthew L. Hedberg,7 Lin Wang,7 Jeffery J.
    [Show full text]
  • Human Induced Pluripotent Stem Cell–Derived Podocytes Mature Into Vascularized Glomeruli Upon Experimental Transplantation
    BASIC RESEARCH www.jasn.org Human Induced Pluripotent Stem Cell–Derived Podocytes Mature into Vascularized Glomeruli upon Experimental Transplantation † Sazia Sharmin,* Atsuhiro Taguchi,* Yusuke Kaku,* Yasuhiro Yoshimura,* Tomoko Ohmori,* ‡ † ‡ Tetsushi Sakuma, Masashi Mukoyama, Takashi Yamamoto, Hidetake Kurihara,§ and | Ryuichi Nishinakamura* *Department of Kidney Development, Institute of Molecular Embryology and Genetics, and †Department of Nephrology, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan; ‡Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Hiroshima, Japan; §Division of Anatomy, Juntendo University School of Medicine, Tokyo, Japan; and |Japan Science and Technology Agency, CREST, Kumamoto, Japan ABSTRACT Glomerular podocytes express proteins, such as nephrin, that constitute the slit diaphragm, thereby contributing to the filtration process in the kidney. Glomerular development has been analyzed mainly in mice, whereas analysis of human kidney development has been minimal because of limited access to embryonic kidneys. We previously reported the induction of three-dimensional primordial glomeruli from human induced pluripotent stem (iPS) cells. Here, using transcription activator–like effector nuclease-mediated homologous recombination, we generated human iPS cell lines that express green fluorescent protein (GFP) in the NPHS1 locus, which encodes nephrin, and we show that GFP expression facilitated accurate visualization of nephrin-positive podocyte formation in
    [Show full text]
  • Rare and Low-Frequency Coding Variants Alter Human Adult Height
    HHS Public Access Author manuscript Author ManuscriptAuthor Manuscript Author Nature. Manuscript Author Author manuscript; Manuscript Author available in PMC 2017 August 01. Published in final edited form as: Nature. 2017 February 09; 542(7640): 186–190. doi:10.1038/nature21039. Rare and low-frequency coding variants alter human adult height A full list of authors and affiliations appears at the end of the article. Summary Height is a highly heritable, classic polygenic trait with ∼700 common associated variants identified so far through genome-wide association studies. Here, we report 83 height-associated coding variants with lower minor allele frequencies (range of 0.1-4.8%) and effects of up to 2 cm/ allele (e.g. in IHH, STC2, AR and CRISPLD2), >10 times the average effect of common variants. In functional follow-up studies, rare height-increasing alleles of STC2 (+1-2 cm/allele) compromised proteolytic inhibition of PAPP-A and increased cleavage of IGFBP-4 in vitro, resulting in higher bioavailability of insulin-like growth factors. These 83 height-associated variants overlap genes mutated in monogenic growth disorders and highlight new biological candidates (e.g. ADAMTS3, IL11RA, NOX4) and pathways (e.g. proteoglycan/ glycosaminoglycan synthesis) involved in growth. Our results demonstrate that sufficiently large sample sizes can uncover rare and low-frequency variants of moderate to large effect associated with polygenic human phenotypes, and that these variants implicate relevant genes and pathways. Reprints and permissions information is available at www.nature.com/reprints.Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http:// www.nature.com/authors/editorial_policies/license.html#terms Correspondence to: Joel N.
    [Show full text]
  • A Homozygous Nonsense Mutation in DCBLD2 Is a Candidate Cause of Developmental Delay, Dysmorphic Features and Restrictive Cardiomyopathy Kheloud M
    www.nature.com/scientificreports OPEN A homozygous nonsense mutation in DCBLD2 is a candidate cause of developmental delay, dysmorphic features and restrictive cardiomyopathy Kheloud M. Alhamoudi1, Tlili Barhoumi2, Hamad Al‑Eidi1, Abdulaziz Asiri3, Marwan Nashabat4, Manal Alaamery5, Masheal Alharbi1, Yazeid Alhaidan1, Brahim Tabarki6, Muhammad Umair1 & Majid Alfadhel1,4* DCBLD2 encodes discodin, CUB and LCCL domain‑containing protein 2, a type‑I transmembrane receptor that is involved in intracellular receptor signalling pathways and the regulation of cell growth. In this report, we describe a 5‑year‑old female who presented severe clinical features, including restrictive cardiomyopathy, developmental delay, spasticity and dysmorphic features. Trio‑ whole‑exome sequencing and segregation analysis were performed to identify the genetic cause of the disease within the family. A novel homozygous nonsense variant in the DCBLD2 gene (c.80G > A, p.W27*) was identifed as the most likely cause of the patient’s phenotype. This nonsense variant falls in the extracellular N‑terminus of DCBLD2 and thus might afect proper protein function of the transmembrane receptor. A number of in vitro investigations were performed on the proband’s skin fbroblasts compared to normal fbroblasts, which allowed a comprehensive assessment resulting in the functional characterization of the identifed DCBLD2 nonsense variant in diferent cellular processes. Our data propose a signifcant association between the identifed variant and the observed reduction in cell proliferation,
    [Show full text]
  • Sun, B. B., Maranville, J. C., Peters, J. E., Stacey, D., Staley, J. R., Blackshaw, J., Burgess, S., Jiang, T., Paige, E., Suren
    Sun, B. B., Maranville, J. C., Peters, J. E., Stacey, D., Staley, J. R., Blackshaw, J., Burgess, S., Jiang, T., Paige, E., Surendran, P., Oliver- Williams, C., Kamat, M. A., Prins, B. P., Wilcox, S. K., Zimmerman, E. S., Chi, A., Bansal, N., Spain, S. L., Wood, A. M., ... Butterworth, A. S. (2018). Genomic atlas of the human plasma proteome. Nature, 558(7708), 73-79. https://doi.org/10.1038/s41586-018-0175-2 Peer reviewed version Link to published version (if available): 10.1038/s41586-018-0175-2 Link to publication record in Explore Bristol Research PDF-document This is the author accepted manuscript (AAM). The final published version (version of record) is available online via Nature Publishing Group at https://www.nature.com/articles/s41586-018-0175-2 . Please refer to any applicable terms of use of the publisher. University of Bristol - Explore Bristol Research General rights This document is made available in accordance with publisher policies. Please cite only the published version using the reference above. Full terms of use are available: http://www.bristol.ac.uk/red/research-policy/pure/user-guides/ebr-terms/ Genomic atlas of the human plasma proteome Benjamin B. Sun1*, Joseph C. Maranville2*, James E. Peters1,3*, David Stacey1, James R. Staley1, James Blackshaw1, Stephen Burgess1,4, Tao Jiang1, Ellie Paige1,5, Praveen Surendran1, Clare Oliver-Williams1,6, Mihir A. Kamat1, Bram P. Prins1, Sheri K. Wilcox7, Erik S. Zimmerman7, An Chi2, Narinder Bansal1,8, Sarah L. Spain9, Angela M. Wood1, Nicholas W. Morrell10, John R. Bradley11, Nebojsa Janjic7, David J. Roberts12,13, Willem H.
    [Show full text]
  • Supplementary Table 1 Double Treatment Vs Single Treatment
    Supplementary table 1 Double treatment vs single treatment Probe ID Symbol Gene name P value Fold change TC0500007292.hg.1 NIM1K NIM1 serine/threonine protein kinase 1.05E-04 5.02 HTA2-neg-47424007_st NA NA 3.44E-03 4.11 HTA2-pos-3475282_st NA NA 3.30E-03 3.24 TC0X00007013.hg.1 MPC1L mitochondrial pyruvate carrier 1-like 5.22E-03 3.21 TC0200010447.hg.1 CASP8 caspase 8, apoptosis-related cysteine peptidase 3.54E-03 2.46 TC0400008390.hg.1 LRIT3 leucine-rich repeat, immunoglobulin-like and transmembrane domains 3 1.86E-03 2.41 TC1700011905.hg.1 DNAH17 dynein, axonemal, heavy chain 17 1.81E-04 2.40 TC0600012064.hg.1 GCM1 glial cells missing homolog 1 (Drosophila) 2.81E-03 2.39 TC0100015789.hg.1 POGZ Transcript Identified by AceView, Entrez Gene ID(s) 23126 3.64E-04 2.38 TC1300010039.hg.1 NEK5 NIMA-related kinase 5 3.39E-03 2.36 TC0900008222.hg.1 STX17 syntaxin 17 1.08E-03 2.29 TC1700012355.hg.1 KRBA2 KRAB-A domain containing 2 5.98E-03 2.28 HTA2-neg-47424044_st NA NA 5.94E-03 2.24 HTA2-neg-47424360_st NA NA 2.12E-03 2.22 TC0800010802.hg.1 C8orf89 chromosome 8 open reading frame 89 6.51E-04 2.20 TC1500010745.hg.1 POLR2M polymerase (RNA) II (DNA directed) polypeptide M 5.19E-03 2.20 TC1500007409.hg.1 GCNT3 glucosaminyl (N-acetyl) transferase 3, mucin type 6.48E-03 2.17 TC2200007132.hg.1 RFPL3 ret finger protein-like 3 5.91E-05 2.17 HTA2-neg-47424024_st NA NA 2.45E-03 2.16 TC0200010474.hg.1 KIAA2012 KIAA2012 5.20E-03 2.16 TC1100007216.hg.1 PRRG4 proline rich Gla (G-carboxyglutamic acid) 4 (transmembrane) 7.43E-03 2.15 TC0400012977.hg.1 SH3D19
    [Show full text]
  • Differentially Expressed Genes in Aneurysm Tissue Compared With
    On-line Table: Differentially expressed genes in aneurysm tissue compared with those in control tissue Fold False Discovery Direction of Gene Entrez Gene Name Function Change P Value Rate (q Value) Expression AADAC Arylacetamide deacetylase Positive regulation of triglyceride 4.46 1.33E-05 2.60E-04 Up-regulated catabolic process ABCA6 ATP-binding cassette, subfamily A (ABC1), Integral component of membrane 3.79 9.15E-14 8.88E-12 Up-regulated member 6 ABCC3 ATP-binding cassette, subfamily C (CFTR/MRP), ATPase activity, coupled to 6.63 1.21E-10 7.33E-09 Up-regulated member 3 transmembrane movement of substances ABI3 ABI family, member 3 Peptidyl-tyrosine phosphorylation 6.47 2.47E-05 4.56E-04 Up-regulated ACKR1 Atypical chemokine receptor 1 (Duffy blood G-protein–coupled receptor signaling 3.80 7.95E-10 4.18E-08 Up-regulated group) pathway ACKR2 Atypical chemokine receptor 2 G-protein–coupled receptor signaling 0.42 3.29E-04 4.41E-03 Down-regulated pathway ACSM1 Acyl-CoA synthetase medium-chain family Energy derivation by oxidation of 9.87 1.70E-08 6.52E-07 Up-regulated member 1 organic compounds ACTC1 Actin, ␣, cardiac muscle 1 Negative regulation of apoptotic 0.30 7.96E-06 1.65E-04 Down-regulated process ACTG2 Actin, ␥2, smooth muscle, enteric Blood microparticle 0.29 1.61E-16 2.36E-14 Down-regulated ADAM33 ADAM domain 33 Integral component of membrane 0.23 9.74E-09 3.95E-07 Down-regulated ADAM8 ADAM domain 8 Positive regulation of tumor necrosis 4.69 2.93E-04 4.01E-03 Up-regulated factor (ligand) superfamily member 11 production ADAMTS18
    [Show full text]
  • DCBLD2 (NM 080927) Human Tagged ORF Clone Product Data
    OriGene Technologies, Inc. 9620 Medical Center Drive, Ste 200 Rockville, MD 20850, US Phone: +1-888-267-4436 [email protected] EU: [email protected] CN: [email protected] Product datasheet for RC224483L4 DCBLD2 (NM_080927) Human Tagged ORF Clone Product data: Product Type: Expression Plasmids Product Name: DCBLD2 (NM_080927) Human Tagged ORF Clone Tag: mGFP Symbol: DCBLD2 Synonyms: CLCP1; ESDN Vector: pLenti-C-mGFP-P2A-Puro (PS100093) E. coli Selection: Chloramphenicol (34 ug/mL) Cell Selection: Puromycin ORF Nucleotide The ORF insert of this clone is exactly the same as(RC224483). Sequence: Restriction Sites: SgfI-MluI Cloning Scheme: ACCN: NM_080927 ORF Size: 2325 bp This product is to be used for laboratory only. Not for diagnostic or therapeutic use. View online » ©2021 OriGene Technologies, Inc., 9620 Medical Center Drive, Ste 200, Rockville, MD 20850, US 1 / 2 DCBLD2 (NM_080927) Human Tagged ORF Clone – RC224483L4 OTI Disclaimer: The molecular sequence of this clone aligns with the gene accession number as a point of reference only. However, individual transcript sequences of the same gene can differ through naturally occurring variations (e.g. polymorphisms), each with its own valid existence. This clone is substantially in agreement with the reference, but a complete review of all prevailing variants is recommended prior to use. More info OTI Annotation: This clone was engineered to express the complete ORF with an expression tag. Expression varies depending on the nature of the gene. RefSeq: NM_080927.3 RefSeq Size: 6093 bp RefSeq ORF: 2328 bp Locus ID: 131566 UniProt ID: Q96PD2 Domains: F5_F8_type_C, CUB, LCCL Protein Families: Druggable Genome, Transmembrane MW: 84.9 kDa This product is to be used for laboratory only.
    [Show full text]
  • Dcbld2/Esdn Is Essential for Proper Optic Tract Formation and Retinal Lamination Ryan Mears Joy University of Vermont
    University of Vermont ScholarWorks @ UVM Graduate College Dissertations and Theses Dissertations and Theses 2016 Dcbld2/esdn Is Essential For Proper Optic Tract Formation And Retinal Lamination Ryan Mears Joy University of Vermont Follow this and additional works at: https://scholarworks.uvm.edu/graddis Part of the Cell and Developmental Biology Commons Recommended Citation Joy, Ryan Mears, "Dcbld2/esdn Is Essential For Proper Optic Tract Formation And Retinal Lamination" (2016). Graduate College Dissertations and Theses. 563. https://scholarworks.uvm.edu/graddis/563 This Thesis is brought to you for free and open access by the Dissertations and Theses at ScholarWorks @ UVM. It has been accepted for inclusion in Graduate College Dissertations and Theses by an authorized administrator of ScholarWorks @ UVM. For more information, please contact [email protected]. DCBLD2/ESDN IS ESSENTIAL FOR PROPER OPTIC TRACT FORMATION AND RETINAL LAMINATION A Thesis Presented by Ryan Joy to The Faculty of the Graduate College of The University of Vermont In Partial Fulfillment of the Requirements for the Degree of Master of Science Specializing in Biology May, 2016 Defense Date: January 7, 2016 Thesis Examination Committee: Bryan A. Ballif, Ph.D., Advisor Jason Stumpff, Ph.D., Chairperson Alicia M. Ebert, Ph.D. Paula Deming, Ph.D., MT Cynthia J. Forehand, Ph.D., Dean of the Graduate College ABSTRACT The Discoidin, CUB and LCCL domain-containing protein 2 (DCBLD2/ESDN/CLCP1) is a type-I, transmembrane receptor that mediates diverse cellular processes such as angiogenesis, vascular remodeling, cellular migration and proliferation. Identification of DCBLD2 in a proteomics screen to identify substrates of Src family tyrosine kinases that bind the Src homology 2 domain of CT10 regulator of kinase-Like (CrkL), a critical scaffolding protein for neuronal development, led to a need for further characterization of the protein.
    [Show full text]
  • Systems Genetics Identifies Sestrin 3 As a Regulator of a Proconvulsant Gene Network in Human Epileptic Hippocampus
    ARTICLE Received 1 Sep 2014 | Accepted 4 Dec 2014 | Published 23 Jan 2015 DOI: 10.1038/ncomms7031 Systems genetics identifies Sestrin 3 as a regulator of a proconvulsant gene network in human epileptic hippocampus Michael R. Johnson1,y, Jacques Behmoaras2,*, Leonardo Bottolo3,*, Michelle L. Krishnan4,*, Katharina Pernhorst5,*, Paola L. Meza Santoscoy6,*, Tiziana Rossetti7, Doug Speed8, Prashant K. Srivastava1,7, Marc Chadeau-Hyam9, Nabil Hajji10, Aleksandra Dabrowska10, Maxime Rotival7, Banafsheh Razzaghi7, Stjepana Kovac11, Klaus Wanisch11, Federico W. Grillo7, Anna Slaviero7, Sarah R. Langley1,7, Kirill Shkura1,7, Paolo Roncon12,13, Tisham De7, Manuel Mattheisen14,15,16, Pitt Niehusmann5, Terence J. O’Brien17, Slave Petrovski18, Marec von Lehe19, Per Hoffmann20,21, Johan Eriksson22,23,24, Alison J. Coffey25, Sven Cichon20,21, Matthew Walker11, Michele Simonato12,13,26,Be´ne´dicte Danis27, Manuela Mazzuferi27, Patrik Foerch27, Susanne Schoch5,28, Vincenzo De Paola7, Rafal M. Kaminski27, Vincent T. Cunliffe6, Albert J. Becker5,y & Enrico Petretto7,29,y Gene-regulatory network analysis is a powerful approach to elucidate the molecular processes and pathways underlying complex disease. Here we employ systems genetics approaches to characterize the genetic regulation of pathophysiological pathways in human temporal lobe epilepsy (TLE). Using surgically acquired hippocampi from 129 TLE patients, we identify a gene-regulatory network genetically associated with epilepsy that contains a specialized, highly expressed transcriptional module encoding proconvulsive cytokines and Toll-like receptor signalling genes. RNA sequencing analysis in a mouse model of TLE using 100 epileptic and 100 control hippocampi shows the proconvulsive module is preserved across-species, specific to the epileptic hippocampus and upregulated in chronic epilepsy.
    [Show full text]