DOCSLIB.ORG

Mouse Il22ra2 Conditional Knockout Project (CRISPR/Cas9)

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Mouse Il22ra2 Conditional Knockout Project (CRISPR/Cas9) https://www.alphaknockout.com Mouse Il22ra2 Conditional Knockout Project (CRISPR/Cas9) Objective: To create a Il22ra2 conditional knockout Mouse model (C57BL/6J) by CRISPR/Cas-mediated genome engineering. Strategy summary: The Il22ra2 gene (NCBI Reference Sequence: NM_178258 ; Ensembl: ENSMUSG00000039760 ) is located on Mouse chromosome 10. 5 exons are identified, with the ATG start codon in exon 1 and the TGA stop codon in exon 5 (Transcript: ENSMUST00000036564). Exon 2 will be selected as conditional knockout region (cKO region). Deletion of this region should result in the loss of function of the Mouse Il22ra2 gene. To engineer the targeting vector, homologous arms and cKO region will be generated by PCR using BAC clone RP23-353K11 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 null mutation display accelerated tumor formation in a colitis associated colon cancer model. Exon 2 starts from about 8.55% of the coding region. The knockout of Exon 2 will result in frameshift of the gene. The size of intron 1 for 5'-loxP site insertion: 2303 bp, and the size of intron 2 for 3'-loxP site insertion: 2092 bp. The size of effective cKO region: ~636 bp. The cKO region does not have any other known gene. Page 1 of 7 https://www.alphaknockout.com Overview of the Targeting Strategy Wildtype allele gRNA region 5' gRNA region 3' 1 2 3 5 Targeting vector Targeted allele Constitutive KO allele (After Cre recombination) Legends Exon of mouse Il22ra2 Homology arm cKO region loxP site Page 2 of 7 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(7136bp) | A(28.71% 2049) | C(21.92% 1564) | T(28.36% 2024) | G(21.01% 1499) 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 7 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% chr10 + 19621152 19624151 3000 browser details YourSeq 100 605 725 3000 95.5% chr14 + 23205578 23205707 130 browser details YourSeq 92 610 735 3000 93.4% chr11 - 63326625 63326807 183 browser details YourSeq 91 610 725 3000 90.2% chr2 - 5103541 5103652 112 browser details YourSeq 91 609 724 3000 93.5% chr15 - 60418442 60418604 163 browser details YourSeq 91 613 720 3000 97.0% chr16 + 40469663 40469932 270 browser details YourSeq 90 610 725 3000 97.0% chr15 + 68499876 68500130 255 browser details YourSeq 89 615 724 3000 96.9% chr1 - 3589217 3589326 110 browser details YourSeq 88 610 725 3000 89.7% chr14 - 118000145 118000256 112 browser details YourSeq 86 616 722 3000 93.2% chr17 - 67845930 67846051 122 browser details YourSeq 85 617 722 3000 96.8% chr11 + 53112227 53112372 146 browser details YourSeq 84 610 725 3000 95.9% chr17 - 16017904 16018053 150 browser details YourSeq 83 609 725 3000 92.0% chr1 + 165284716 165284844 129 browser details YourSeq 81 614 724 3000 90.4% chr2 - 80452519 80452637 119 browser details YourSeq 77 610 690 3000 98.8% chr13 - 76926781 76926906 126 browser details YourSeq 76 616 706 3000 96.3% chr14 - 25264168 25264257 90 browser details YourSeq 76 610 710 3000 90.3% chr13 - 114992966 114993061 96 browser details YourSeq 76 610 725 3000 88.8% chr10 - 120964321 120964423 103 browser details YourSeq 76 611 722 3000 88.6% chr11 + 48417919 48418062 144 browser details YourSeq 71 627 724 3000 91.2% chr16 + 26065417 26065512 96 Note: The 3000 bp section upstream of Exon 2 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% chr10 + 19624788 19627787 3000 browser details YourSeq 42 2546 2588 3000 100.0% chr16 - 22475031 22475074 44 browser details YourSeq 42 2546 2587 3000 100.0% chr17 + 43398336 43398377 42 browser details YourSeq 41 2546 2586 3000 100.0% chr13 - 46057342 46057382 41 browser details YourSeq 41 2546 2586 3000 100.0% chr12 - 21557709 21557749 41 browser details YourSeq 41 2546 2586 3000 100.0% chr3 + 50272583 50272623 41 browser details YourSeq 41 2546 2586 3000 100.0% chr13 + 95612750 95612790 41 browser details YourSeq 41 2546 2586 3000 100.0% chr12 + 13213057 13213097 41 browser details YourSeq 40 2546 2585 3000 100.0% chr14 - 106606351 106606390 40 browser details YourSeq 40 2545 2584 3000 100.0% chr12 - 29145202 29145241 40 browser details YourSeq 40 2546 2585 3000 100.0% chr11 - 102003133 102003172 40 browser details YourSeq 39 2545 2584 3000 100.0% chr10 - 92262168 92262211 44 browser details YourSeq 39 2548 2586 3000 100.0% chr18 + 38295946 38295984 39 browser details YourSeq 38 2548 2585 3000 100.0% chr3 + 157643096 157643133 38 Note: The 3000 bp section downstream of Exon 2 is BLAT searched against the genome. No significant similarity is found. Page 4 of 7 https://www.alphaknockout.com Gene and protein information: Il22ra2 interleukin 22 receptor, alpha 2 [ Mus musculus (house mouse) ] Gene ID: 237310, updated on 1-Oct-2019 Gene summary Official Symbol Il22ra2 provided by MGI Official Full Name interleukin 22 receptor, alpha 2 provided by MGI Primary source MGI:MGI:2665114 See related Ensembl:ENSMUSG00000039760 Gene type protein coding RefSeq status PROVISIONAL Organism Mus musculus Lineage Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia; Eutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae; Murinae; Mus; Mus Also known as CRF2X; CRF2-10; CRF2-s1; Il-22bp Expression Low expression observed in reference dataset See more Orthologs human all Genomic context Location: 10; 10 A3 See Il22ra2 in Genome Data Viewer Exon count: 5 Annotation release Status Assembly Chr Location 108 current GRCm38.p6 (GCF_000001635.26) 10 NC_000076.6 (19622028..19634681) Build 37.2 previous assembly MGSCv37 (GCF_000001635.18) 10 NC_000076.5 (19341834..19354487) Chromosome 10 - NC_000076.6 Page 5 of 7 https://www.alphaknockout.com Transcript information: This gene has 1 transcript Gene: Il22ra2 ENSMUSG00000039760 Description interleukin 22 receptor, alpha 2 [Source:MGI Symbol;Acc:MGI:2665114] Gene Synonyms Il-22bp Location Chromosome 10: 19,621,998-19,634,681 forward strand. GRCm38:CM001003.2 About this gene This gene has 1 transcript (splice variant), 137 orthologues, 12 paralogues, is a member of 1 Ensembl protein family and is associated with 4 phenotypes. Transcripts Name Transcript ID bp Protein Translation ID Biotype CCDS UniProt Flags Il22ra2-201 ENSMUST00000036564.7 2498 230aa ENSMUSP00000042642.7 Protein coding CCDS23717 Q80XF5 TSL:1 GENCODE basic APPRIS P1 32.68 kb Forward strand 19.62Mb 19.63Mb 19.64Mb Genes (Comprehensive set... Il22ra2-201 >protein coding Contigs AC153862.8 > Regulatory Build 19.62Mb 19.63Mb 19.64Mb Reverse strand 32.68 kb Regulation Legend CTCF Enhancer Open Chromatin Promoter Flank Transcription Factor Binding Site Gene Legend Protein Coding merged Ensembl/Havana Page 6 of 7 https://www.alphaknockout.com Transcript: ENSMUST00000036564 12.68 kb Forward strand Il22ra2-201 >protein coding ENSMUSP00000042... Low complexity (Seg) Cleavage site (Sign... Superfamily Fibronectin type III superfamily Pfam Fibronectin type III Interferon/interleukin receptor domain PROSITE profiles Fibronectin type III PANTHER PTHR20859 PTHR20859:SF51 Gene3D Immunoglobulin-like fold CDD Fibronectin type III All sequence SNPs/i... Sequence variants (dbSNP and all other sources) Variant Legend missense variant synonymous variant Scale bar 0 20 40 60 80 100 120 140 160 180 200 230 We wish to acknowledge the following valuable scientific information resources: Ensembl, MGI, NCBI, UCSC. Page 7 of 7.
Load more

Recommended publications
  • Cytokine Nomenclature
    RayBiotech, Inc. The protein array pioneer company Cytokine Nomenclature Cytokine Name Official Full Name Genbank Related Names Symbol 4-1BB TNFRSF Tumor necrosis factor NP_001552 CD137, ILA, 4-1BB ligand receptor 9 receptor superfamily .2. member 9 6Ckine CCL21 6-Cysteine Chemokine NM_002989 Small-inducible cytokine A21, Beta chemokine exodus-2, Secondary lymphoid-tissue chemokine, SLC, SCYA21 ACE ACE Angiotensin-converting NP_000780 CD143, DCP, DCP1 enzyme .1. NP_690043 .1. ACE-2 ACE2 Angiotensin-converting NP_068576 ACE-related carboxypeptidase, enzyme 2 .1 Angiotensin-converting enzyme homolog ACTH ACTH Adrenocorticotropic NP_000930 POMC, Pro-opiomelanocortin, hormone .1. Corticotropin-lipotropin, NPP, NP_001030 Melanotropin gamma, Gamma- 333.1 MSH, Potential peptide, Corticotropin, Melanotropin alpha, Alpha-MSH, Corticotropin-like intermediary peptide, CLIP, Lipotropin beta, Beta-LPH, Lipotropin gamma, Gamma-LPH, Melanotropin beta, Beta-MSH, Beta-endorphin, Met-enkephalin ACTHR ACTHR Adrenocorticotropic NP_000520 Melanocortin receptor 2, MC2-R hormone receptor .1 Activin A INHBA Activin A NM_002192 Activin beta-A chain, Erythroid differentiation protein, EDF, INHBA Activin B INHBB Activin B NM_002193 Inhibin beta B chain, Activin beta-B chain Activin C INHBC Activin C NM005538 Inhibin, beta C Activin RIA ACVR1 Activin receptor type-1 NM_001105 Activin receptor type I, ACTR-I, Serine/threonine-protein kinase receptor R1, SKR1, Activin receptor-like kinase 2, ALK-2, TGF-B superfamily receptor type I, TSR-I, ACVRLK2 Activin RIB ACVR1B
    [Show full text]
  • IL-21R, Human, Recombinant (Sf9) Recombinant Human Interleukin 21 Receptor (Sf9 Cell-Derived)
    IL-21R, human, recombinant (Sf9) Recombinant Human Interleukin 21 Receptor (Sf9 cell-derived) Instruction Manual Catalog Number C-62924 Synonyms Interleukin 21 Receptor, Novel Interleukin Receptor, IL-21 Receptor, NILR, Interleukin-21 Receptor, CD360 Antigen, IL-21R, CD360 Description Interleukin-21 receptor, also known as IL-21R is a member of the type I cytokine receptors family. IL-21R forms a heterodimeric receptor complex with the common gamma-chain, a receptor subunit which is also shared by the receptors for Interleukin 2, 4, 7, 9, and 15. Furthermore, IL-21 receptor transduces the growth promoting signal of IL-21, and is significant for the proliferation as well as differentiation of T cells, B cells, and natural killer (NK) cells. The ligand binding of IL-21 receptor leads to the activation of numerous downstream signaling molecules, including JAK1, JAK3, STAT1, as well as STAT3. IL21R produced in Sf9 cells is a single, glycosylated polypeptide chain (20-232 a.a.) fused to an 8 aa His-tag at the C-terminus. It contains a total of 221 amino acids and has a molecular mass of 25.6 kDa. IL-21R shows multiple bands between 28-40 kDa on SDS-PAGE under reducing conditions and has been purified using proprietary chromatographic techniques. Quantity 10 µg Molecular Mass 25.6 kDa Source Sf9 cells Biological-Activity NA Specific Activity NA Formulation Sterile-filtered colorless protein solution (1 mg/ml) containing phosphate buffered saline (pH 7.4) and 10% glycerol. Reconstitution Please Note: Always centrifuge product briefly before opening vial. The dissolved protein can be diluted into other aqueous buffers and stored at -20°C for future use.
    [Show full text]
  • Structure-Based Tuning of Interleukin Receptor Complexes to Promote Anti- Tumor Immunity
    Science Highlight – July 2021 Structure-based Tuning of Interleukin Receptor Complexes to Promote Anti- tumor Immunity Cells of the immune system communicate messages between them via small secreted complexes called cytokines. Cytokines are recognized by other cells through membrane receptors. These cell signaling pathways convey information about pathogens, cancers, or other problems that concern the immune system. Two of these cytokines, interleukin 12 (IL- 12) and Interleukin 23 (IL-23) are made by antigen-presenting cells and help to activate lymphocytes, including both T and NK cells. IL-12 and IL-23 have similar structures, which include a four-helix bundle α-subunit and a β- subunit called p40. Despite intense basic research and clinical interest in IL-12 and IL-23, a structural basis for receptor assembly has remained elusive. In work led by the Garcia group at Stanford University, researchers determined the crystal structure of the complete IL-23 receptor complex using x-ray crystallography data collected on SSRL beam line 12-2. Figure 1. Structural mechanism of IL-12 and IL-23 receptor assembly enables graded control of downstream signaling. (A) The 3.4 Å resolution crystal structure of the IL-23 receptor complex. P40 (orange) bridges IL-23 and IL-12. (B-C) CryoEM maps of the IL-23 (8 Å) and IL-12 (10 Å) receptor complexes, respectively. (D-E) Graded control of IL-12 and IL-23 signaling. STAT phosphorylation was analyzed by flow cytometry. The structure revealed that IL-23 uses the four-helix bundle and p40 subunits to engage its’ receptors in a modular fashion (Fig.
    [Show full text]
  • Anti-Human IL-21 Purified Catalog Number: 14-6465 Also Known As: Interleukin-21, IL21 RUO: for Research Use Only
    Page 1 of 2 Anti-Human IL-21 Purified Catalog Number: 14-6465 Also known as: Interleukin-21, IL21 RUO: For Research Use Only. Not for use in diagnostic procedures. Immunoblot analysis of reduced HL60 cell lysates using Anti-Human IL-21 Purified (1µg/ml) and detected using Anti-Rabbit IgG-HRP. Product Information Contents: Anti-Human IL-21 Purified Formulation: aqueous buffer, 0.09% sodium Catalog Number: 14-6465 azide, may contain carrier protein/stabilizer Clone: Polyclonal Temperature Limitation: Store at 2-8°C. Host/Isotype: Rabbit IgG Batch Code: Refer to vial Use By: Refer to vial Caution, contains Azide Description The rabbit polyclonal antibody reacts with human IL-21; the antibody was raised against a synthetic peptide (tcpscdsyekkppke) corresponding to amino acids 121 to 135 of human IL-21 precursor (1). A novel cytokine was recently identified in human and mouse and designated IL-21 (1), which has significant homology to IL-2, IL-4, and IL- 15. The receptor for IL-21 (IL-21R, also termed NILR for novel Interleukin receptor) is a new member of the class I cytokine receptor family (1,2). IL-21R forms a complex with the common cytokine receptor g chain, gc, and mediates IL-21 signaling (3,4). IL-21 and its receptor activate JAK-STAT signaling pathway. IL-21 is expressed in activated T cells, and HL-60 and THP-1 cell lines. IL-21 plays a role in the proliferation and maturation of NK, B and T cell populations. Applications Reported This polyclonal antibody has been reported for use in immunoblotting (WB).
    [Show full text]
  • Human IL-18R1 Accusignal ELISA Kit - KOA0744
    Human IL-18R1 AccuSignal ELISA Kit - KOA0744 Code: KOA0744 Size: 1 Kit Product Description: Human IL-18R1 AccuSignal ELISA Kit - KOA0744 PhysicalState: Label Unconjugated Gene Name IL18R1 Species Reactivity Human Storage Condition Store vials at 4°C prior to opening. Centrifuge product if not completely clear after standing at room temperature. This product is stable for 6 months at 4°C as an undiluted liquid. Dilute only prior to immediate use. For extended storage freeze at -20°C or below for 12 months. Avoid cycles of freezing and thawing. Synonyms CD218 antigen-like family member A, CD218a, CDw218a, CDw218a antigen, IL 1Rrp, IL-18R-1, IL-18R1, IL- 1Rrp, IL1 receptor related protein, IL1 receptor-related protein, IL18R_HUMAN, IL18R1, IL18RA, Il18ralpha, IL1R-rp, IL1RRP, Interleukin 18 receptor 1, Interleukin 18 receptor alpha chain, Interleukin-18 receptor 1 Application Note Useful in Sandwich ELISA for Quantitative Detection of Antigen. Aliquot 0.1ml per well of the 2000pg/ml, 1000pg/ml, 500pg/ml, 250pg/ml, 125pg/ml, 62.5pg/ml, 31.2pg/ml human IL-18R1 standard solutions into the precoated 96-well plate. Add 0.1ml of the sample diluent buffer into the control well (Zero well). Add 0.1ml of each properly diluted sample of human cell culture supernates, serum or plasma(heparin, EDTA) to each empty well. It is recommended that each human IL-18R1 standard solution and each sample be measured in duplicate. Background The interleukin-18 receptor 1(IL-18R1), also known as CDw218a(cluster of differentiation w218a) or IL18RA, is an interleukin receptor of the immunoglobulin superfamily.
    [Show full text]
  • The Interleukin-18 Receptor Is Differentially Expressed in Whole
    1 The interleukin-18 receptor is differentially expressed in the blood during sepsis. 2 3 Shahan Mamoor, MS1 4 [email protected] East Islip, NY, USA 5 6 We probed published and public microarray datasets1,2 to discover the most significant gene expression changes in the blood of patients with sepsis. We found significant induction 7 expression of IL18RAP and IL18R1, genes encoding subunits of the interleukin-18 receptor, in 8 whole blood from patients with sepsis. 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 Keywords: sepsis, septic shock, interleukin-18, IL18R1, IL18RAP, systems biology of septic 26 shock. 27 28 PAGE 1 OF 13 1 Septic shock is a leading cause of mortality in the United States and worldwide3. We 2 used published and public microarray datasets1,2 to identify differentially expressed genes in the 3 4 blood of patients with sepsis. We identified IL18R1 and IL18RAP as among the genes most 5 differentially expressed in blood in the septic state. 6 7 Methods 8 9 We utilized microarray datasets GSE1001591 and GSE264402 for this differential gene 10 11 expression analysis of blood cells during sepsis. GSE100509 was generated with whole blood 12 using Illumina HumanWG-6 v3.0 expression beadchip technology with n=12 whole blood from 13 control subjects and n=33 whole blood from sepsis patients. GSE26440 was generated using 14 15 Affymetrix Human Genome U133 Plus 2.0 Array technology with n=32 control subjects and 16 n=98 sepsis patients. The Benjamini and Hochberg method of p-value adjustment was used for 17 ranking of differential expression but raw p-values were used for assessment of statistical 18 19 significance of global differential expression.
    [Show full text]
  • Open Chen-Thesis Finalv5.Pdf
    The Pennsylvania State University The Graduate School Department of Neural and Behavioral Sciences EPIGENETIC ANALYSIS OF IMMUNE ASSOCIATED SIGNALING MOLECULES DURING MOUSE RETINA DEVELOPMENT A Thesis in Anatomy by Chen Yang © 2013 Chen Yang Submitted in Partial Fulfillment of the Requirements for the Degree of Master of Science May 2013 The thesis of Chen Yang was reviewed and approved* by the following: Samuel Shao-Min Zhang Assistant Professor of Neural and Behavioral Sciences Thesis Advisor Colin J. Barnstable Department Head of Neural and Behavioral Sciences Professor of Neural and Behavioral Sciences Patricia J. McLaughlin Professor of Neural and Behavioral Sciences Director of Graduate Program in Anatomy *Signatures are on file in the Graduate School. ii ABSTRACT The retina is an immune-privileged organ. Many autoimmune diseases, such as AMD, glaucoma, and diabetic retinopathy, are caused by excessive inflammatory responses targeting self-tissue. The physiological functions of extracellular and intracellular signaling molecules of immune responses have been well characterized. The epigenetic aspects of these molecules in the retina, however, have not been well elucidated. In this study, we examined the expression of selected immune-related genes, and their transcriptional accessibility via epigenetic mapping, cluster analysis, and RT-PCR. Among these genes, interleukin receptor related genes and intracellular signaling molecules exhibit higher transcriptional accessibility. Epigenetic mapping of the toll-like receptor (TLR) family revealed that 3 out of 13 TLRs exhibit H3K4me2 accumulation during retina development, suggesting that TLR2, TLR3, and TLR9 are the only TLR members expressed in the retina. Most of the NF-κB signaling molecules exhibited transcriptional accessibility, implying their essential roles in inflammatory regulation during retina maturation.
    [Show full text]
  • 32-6465: IL18R1 Human Description Product Info
    9853 Pacific Heights Blvd. Suite D. San Diego, CA 92121, USA Tel: 858-263-4982 Email: [email protected] 32-6465: IL18R1 Human Interleukin 18 Receptor 1, CD218 Antigen-Like Family Member A, IL1 Receptor-Related Protein, IL-18R-1, Alternative Name CDw218a, IL-18R1, IL-1Rrp, IL1R-Rp, IL1RRP, Interleukin-18 Receptor 1, Cytokine Receptor, CD218a : Antigen, IL18Ralpha2, CD218a, IL18RA. Description Source: Sf9, Baculovirus cells. Sterile filtered colorless solution. IL18R1 (interleukin 18 receptor 1) is an interleukin receptor which is part of the immunoglobulin superfamily. IL18R1 comprises 3 Ig-like C2-type domains, on TIR domain IL18R1 shares many immuno-regulatory functions with IL12. IL12 and IFN-alpha are found to induce the expression of IL18 receptor in NK & T cells. Moreover, IL18R1 explicitly binds interleukin 18, and is vital for IL18 mediated signal transduction. IL18R1 does not bind IL1A or IL1B beta, however it does bind to the agonist which causes NF-kappa-B to become active. IL18R1 produced in Sf9 Baculovirus cells is a single, glycosylated polypeptide chain containing a total of 550 amino acids (19-329 a.a.) and having a molecular mass of 62.7kDa (Molecular size on SDS-PAGE will appear at approximately 70-100kDa). IL18R1 is expressed with a 239 amino acid hIgG-His tag at C-Terminus and purified by proprietary chromatographic techniques. Product Info Amount : 2 µg / 10 µg Purification : Greater than 90.0% as determined by SDS-PAGE. Content : IL18R1 protein solution (0.5mg/ml) contains 10% glycerol & Phosphate buffered saline (pH7.4). Store at 4°C if entire vial will be used within 2-4 weeks.
    [Show full text]
  • Cynomolgus IL-21R / Interleukin-21 Receptor Protein (His Tag)
    Cynomolgus IL-21R / Interleukin-21 Receptor Protein (His Tag) Catalog Number: 90315-C08H General Information SDS-PAGE: Gene Name Synonym: IL21R Protein Construction: A DNA sequence encoding the cynomolgus IL21R (XP_005591578.1) (Met1-Pro236) was expressed with a polyhistidine tag at the C-terminus. Source: Cynomolgus Expression Host: HEK293 Cells QC Testing Purity: > 90 % as determined by SDS-PAGE Endotoxin: Protein Description < 1.0 EU per μg of the protein as determined by the LAL method Interleukin-21 receptor, also known as IL-21 receptor, IL-21R, Novel Stability: interleukin receptor, IL21R and NILR, is a single-pass type I membrane protein which belongs to the type I cytokine receptor family and Type 4 Samples are stable for up to twelve months from date of receipt at -70 ℃ subfamily. Interleukin-21 (IL-21) belongs to a family of cytokines that bind to a composite receptor consisting of a private receptor (IL-21R) and the Predicted N terminal: Cys 20 common cytokine receptor gamma chain ( gamma(C) ). The IL-21R is Molecular Mass: discovered as a novel member of the class-I-cytokine-receptor family and is selectively expressed in lymphoid tissues. IL-21R shows strong The recombinant cynomolgus IL21R comprises 228 amino acids and has a sequence homologies to the interleukin-4 receptor alpha chain gene (IL- calculated molecular mass of 26.4 KDa. The apparent molecular mass of it 4RA). The WSXWS motif of IL-21R appears to be necessary for proper is approximately 44.5 KDa in SDS-PAGE. protein folding and thereby efficient intracellular transport and cell-surface receptor binding.
    [Show full text]
  • Identification and Characterization of Multiple Splice Forms of the Human Interleukin-23 Receptor a Chain in Mitogen-Activated Leukocytes
    Genes and Immunity (2008) 9, 631–639 & 2008 Macmillan Publishers Limited All rights reserved 1466-4879/08 $32.00 www.nature.com/gene ORIGINAL ARTICLE Identification and characterization of multiple splice forms of the human interleukin-23 receptor a chain in mitogen-activated leukocytes S-h Kan, G Mancini and G Gallagher Genetic Immunology Laboratory, HUMIGEN LLC, The Institute for Genetic Immunology, Hamilton, NJ, USA The signalling of interleukin-23 (IL-23) and its receptor (IL-23R) is a key element in the differentiation of T cells to the Th17 phenotype. Here, we present the identification and characterization of human IL23R splice variants resulting from alternative splicing of the IL23R mRNA, from activated human leukocytes, following the analysis of IL23R cDNA. Twenty-four different IL23R transcripts were observed in this study, which may potentially lead to an alteration in the protein coding region of IL-23Ra. Consequently, by analysing amino acid sequences deduced from alternatively spliced mRNA sequences, four different putative premature early termination forms of IL-23Ra: (1) a very short ‘IL-23Ra’, (2) an IL-23Ra containing only the extracellular region, (3) a IL-23Ra with truncated intracellular domain and (4) in-frame exon-skipping causing changes to the extracellular region of the IL-23Ra were revealed. These changes may affect the function of IL-23R by altering the ligand–binding interaction, producing a soluble form of the receptor to act as a decoy receptor and/or modify the IL-23/IL-23R signalling, respectively. Taken together, identification of potentially functional splice variants of IL23R underscores the biological diversity of the IL23R gene and will aid in the understanding of the gene’s function in normal and pathological conditions.
    [Show full text]
  • Inherited Human IRAK-1 Deficiency Selectively Impairs TLR Signaling in Fibroblasts
    Inherited human IRAK-1 deficiency selectively impairs TLR signaling in fibroblasts Erika Della Minaa,b, Alessandro Borghesic,d, Hao Zhoue,1, Salim Bougarnf,1, Sabri Boughorbelf,1, Laura Israela,b, Ilaria Melonig, Maya Chrabieha,b, Yun Linga,b, Yuval Itanh, Alessandra Renierig,i, Iolanda Mazzucchellid,j, Sabrina Bassok, Piero Pavonel, Raffaele Falsaperlal, Roberto Cicconem, Rosa Maria Cerboc, Mauro Stronatic,d, Capucine Picarda,b,n,o, Orsetta Zuffardim, Laurent Abela,b,h, Damien Chaussabelf,2, Nico Marrf,2, Xiaoxia Lie,2, Jean-Laurent Casanovaa,b,h,n,p,3,4, and Anne Puela,b,h,3,4 aLaboratory of Human Genetics of Infectious Diseases, Necker Branch, INSERM U1163, 75015 Paris, France; bImagine Institute, Paris Descartes University, 75015 Paris, France; cNeonatal Intensive Care Unit, Instituto di Ricovero e Cura a Carattere Scientifico (IRCCS) San Matteo Hospital Foundation, 27100 Pavia, Italy; dLaboratory of Neonatal Immunology, IRCCS San Matteo Hospital Foundation, 27100 Pavia, Italy; eDepartment of Immunology, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH 44106; fSidra Medical and Research Center, Doha, Qatar; gMedical Genetics, Department of Medical Biotechnologies, University of Siena, 53100 Siena, Italy; hSt. Giles Laboratory of Human Genetics of Infectious Diseases, Rockefeller Branch, The Rockefeller University, New York, NY 10065; iMedical Genetics, University Hospital of Siena, 53100 Siena, Italy; jDepartment of Internal Medicine and Therapeutics, University of Pavia, 27100 Pavia, Italy; kLaboratory of Transplant
    [Show full text]
  • Integrating Mendelian Randomization and Multiple-Trait Colocalization to Uncover 2 Cell-Specific Inflammatory Drivers of Autoimmune and Atopic Disease
    bioRxiv preprint doi: https://doi.org/10.1101/394296; this version posted August 17, 2018. 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 Integrating Mendelian randomization and multiple-trait colocalization to uncover 2 cell-specific inflammatory drivers of autoimmune and atopic disease 3 Lucy M. McGowan 1,*, George Davey Smith2, Tom R. Gaunt2, Tom G. Richardson2,* 4 Abstract 5 Immune mediated diseases (IMDs) arise from a lack of immune tolerance, causing chronic 6 inflammation. Despite their growing prevalence, targeted therapies to treat IMDs are lacking. 7 Cytokines and their receptors, which mediate inflammation, have been associated with IMD 8 susceptibility. However, the complex signalling networks and multiple cell-types required to 9 orchestrate inflammatory responses have made it difficult to pinpoint specific cytokines and immune 10 cell-types which drive IMDs. 11 In this study, we developed an analytical framework which integrates Mendelian randomisation (MR) 12 and multiple-trait colocalization (moloc) analyses to determine putative cell-specific drivers of IMDs. 13 We used MR to determine the likelihood of causal associations between the levels of 10 circulating 14 cytokines/cytokine receptors and 9 IMDs within human populations of European descent. 15 Conservative (single SNP) and liberal (multiple SNP) MR analysis supported a causal role for IL-18 in 16 inflammatory bowel disease (P = 1.17 x 10-4) and eczema/dermatitis (P = 2.81 x 10-3), as well as roles 17 for IL-2rα and IL-6R in several IMDs.
    [Show full text]