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The Cell Engineering Facility uses state of the art CRISPR/Cas techniques to generate gene knock-outs and knock-ins in target cells. Other applications of CRISPR/Cas may be possible following consultation.
Gene knock-out means that the function of a specific gene, and consequently that of its encoded protein, is disrupted by a specific mutation. Gene knock-in means that the specific target gene and its protein gain a new function because of an engineered mutation. By using the resulting CRISPR/Cas cell clones, the function of specific genes and proteins can be studied in detail within the cellular context.
After its publication in 2012, the CRISPR/Cas gene editing system revolutionized molecular biology within only a few years (CRISPR = clustered regularly interspaced short palindromic repeats; see also our Introduction page). The increase in publications using CRISPR/Cas is only comparable to what was seen in the eighties following the development of the polymerase chain reaction (PCR) or in the nineties following the introduction of the green fluorescent protein (GFP). The Cell Engineering Facility of the FMP will make gene knock-outs and knock-ins in various target cells using state-of-the art CRISPR/Cas techniques. Other applications may be possible following consultation. Cell clones will be characterized by DNA sequencing and the genotype will be analyzed to confirm modification of both alleles. Western blotting will be done in case you could provide a specific antibody.
An overview of the methodology we use can be found on the “Methods” page. If you are already familiar with CRISPR/Cas, you may immediately proceed to the “Order” page. Depending on the amount of simultaneous orders, clones should be ready within 8-12 weeks depending on cell type. If you use the system for the first time or need an overview of its history, function and applications, you may visit our “Introduction” page first. If you have any questions, please get in touch.
In agreement with the FMP directors, we will charge will charge 1,000 Euro € for a CRISPR/Cas cell clone (knock-in or knock out) when the clone is confirmed.
In addition to CRISPR/Cas gene editing, the Cell Engineering Facility is also able to make stable transfected cell clones using various cell lines upon request.
Inhibitors of the Sec61 Complex and Novel High Throughput Screening Strategies to Target the Protein Translocation Pathway
International journal of molecular sciences 2021
read onlineA Proteomic Study on the Membrane Protein Fraction of T Cells Confirms High Substrate Selectivity for the ER Translocation Inhibitor Cyclotriazadisulfonamide
Molecular & cellular proteomics : MCP 2021
read onlineModulating TSH Receptor Signaling for Therapeutic Benefit
European thyroid journal 2020
read onlineCharacterization of five novel vasopressin V2 receptor mutants causing nephrogenic diabetes insipidus reveals a role of tolvaptan for M272R-V2R mutation
Scientific reports 2020
read onlineA New Highly Thyrotropin Receptor-Selective Small-Molecule Antagonist with Potential for the Treatment of Graves' Orbitopathy
Thyroid : official journal of the American Thyroid Association 2019
read onlineThyrotropin Receptor: Allosteric Modulators Illuminate Intramolecular Signaling Mechanisms at the Interface of Ecto- and Transmembrane Domain
Molecular pharmacology 2019
read onlineEvidence of G-protein-coupled receptor and substrate transporter heteromerization at a single molecule level
Cellular and molecular life sciences : CMLS 2018
read onlineUse of a sequential high throughput screening assay to identify novel inhibitors of the eukaryotic SRP-Sec61 targeting/translocation pathway
PloS one 2018
read onlineEvidence for Heterodimerization and Functional Interaction of the Angiotensin Type 2 Receptor and the Receptor MAS
Hypertension (Dallas, Tex. : 1979) 2017
read onlineTargeting G Protein-Coupled Receptors by Capture Compound Mass Spectrometry: A Case Study with Sertindole
Chembiochem : a European journal of chemical biology 2017
read onlineFunctional Significance of the Signal Peptides of Corticotropin-Releasing Factor Receptors
Current molecular pharmacology 2017
read onlineDefining a conformational consensus motif in cotransin-sensitive signal sequences: a proteomic and site-directed mutagenesis study
PloS one 2015
read onlineA new variant in signal peptide of the human luteinizing hormone receptor (LHCGR) affects receptor biogenesis causing leydig cell hypoplasia
Human molecular genetics 2015
read onlineInvolvement of the L-Type Amino Acid Transporter Lat2 in the Transport of 3,3'-Diiodothyronine across the Plasma Membrane
European thyroid journal 2015
read onlineN-Terminal Signal Peptides of G Protein-Coupled Receptors: Significance for Receptor Biosynthesis, Trafficking, and Signal Transduction
Progress in molecular biology and translational science 2015
read onlineStructural insights into thyroid hormone transport mechanisms of the L-type amino acid transporter 2
Molecular endocrinology (Baltimore, Md.) 2015
read onlineThe specific monomer/dimer equilibrium of the corticotropin-releasing factor receptor type 1 is established in the endoplasmic reticulum
The Journal of biological chemistry 2014
read onlineDifferences between lutropin-mediated and choriogonadotropin-mediated receptor activation
The FEBS journal 2014
read onlineUse of Kaede and Kikume green-red fusions for live cell imaging of G protein-coupled receptors
Methods in molecular biology (Clifton, N.J.) 2014
read onlineThe Pseudo signal peptide of the corticotropin-releasing factor receptor type 2A prevents receptor oligomerization
The Journal of biological chemistry 2012
read onlineThe protease-activated receptor 1 possesses a functional and cleavable signal peptide which is necessary for receptor expression
FEBS letters 2012
read onlineLive cell imaging of G protein-coupled receptors
Methods in molecular biology (Clifton, N.J.) 2012
read onlineUse of Kikume green-red fusions to study the influence of pharmacological chaperones on trafficking of G protein-coupled receptors
FEBS letters 2012
read onlineFunctional significance of cleavable signal peptides of G protein-coupled receptors
European journal of cell biology 2012
read onlineThe proteoglycan syndecan 4 regulates transient receptor potential canonical 6 channels via RhoA/Rho-associated protein kinase signaling
Arteriosclerosis, thrombosis, and vascular biology 2012
read onlineInhibition of biosynthesis of human endothelin B receptor by the cyclodepsipeptide cotransin
The Journal of biological chemistry 2011
read onlineMutations that silence constitutive signaling activity in the allosteric ligand-binding site of the thyrotropin receptor
Cellular and molecular life sciences : CMLS 2011
read onlineFrom molecular details of the interplay between transmembrane helices of the thyrotropin receptor to general aspects of signal transduction in family a G-protein-coupled receptors (GPCRs)
The Journal of biological chemistry 2011
read onlinePrimary and secondary thyroid hormone transporters
Thyroid research 2011
read onlineThe pseudo signal peptide of the corticotropin-releasing factor receptor type 2a decreases receptor expression and prevents Gi-mediated inhibition of adenylyl cyclase activity
The Journal of biological chemistry 2010
read onlineSignaling-sensitive amino acids surround the allosteric ligand binding site of the thyrotropin receptor
FASEB journal : official publication of the Federation of American Societies for Experimental Biology 2010
read onlineUse of Kaede fusions to visualize recycling of G protein-coupled receptors
Traffic (Copenhagen, Denmark) 2009
read onlineThe sequence after the signal peptide of the G protein-coupled endothelin B receptor is required for efficient translocon gating at the endoplasmic reticulum membrane
Molecular pharmacology 2009
read onlineThe corticotropin-releasing factor receptor type 2a contains an N-terminal pseudo signal peptide
The Journal of biological chemistry 2006
read onlineTIRC7 inhibits T cell proliferation by modulation of CTLA-4 expression
Journal of immunology (Baltimore, Md. : 1950) 2006
read onlineThe signal peptide of the rat corticotropin-releasing factor receptor 1 promotes receptor expression but is not essential for establishing a functional receptor
The Biochemical journal 2005
read onlineThe role of N-glycosylation in the stability, trafficking and GABA-uptake of GABA-transporter 1. Terminal N-glycans facilitate efficient GABA-uptake activity of the GABA transporter
The FEBS journal 2005
read onlineThe hydrophobic amino acid residues in the membrane-proximal C tail of the G protein-coupled vasopressin V2 receptor are necessary for transport-competent receptor folding
FEBS letters 2005
read onlineA Ser/Thr cluster within the C-terminal domain of the rat prostaglandin receptor EP3alpha is essential for agonist-induced phosphorylation, desensitization and internalization
British journal of pharmacology 2005
read onlineIdentification of a Ser/Thr cluster in the C-terminal domain of the human prostaglandin receptor EP4 that is essential for agonist-induced beta-arrestin1 recruitment but differs from the apparent principal phosphorylation site
The Biochemical journal 2004
read onlineThe early stages of the intracellular transport of membrane proteins: clinical and pharmacological implications
Reviews of physiology, biochemistry and pharmacology 2004
read onlinePharmacochaperones post-translationally enhance cell surface expression by increasing conformational stability of wild-type and mutant vasopressin V2 receptors
The Journal of biological chemistry 2004
read onlineDisease-causing V(2) vasopressin receptors are retained in different compartments of the early secretory pathway
Traffic (Copenhagen, Denmark) 2004
read onlineThe extracellular N terminus of the endothelin B (ETB) receptor is cleaved by a metalloprotease in an agonist-dependent process
The Journal of biological chemistry 2002
read onlineThe signal peptide of the G protein-coupled human endothelin B receptor is necessary for translocation of the N-terminal tail across the endoplasmic reticulum membrane
The Journal of biological chemistry 2002
read onlineFunctional rescue of the nephrogenic diabetes insipidus-causing vasopressin V2 receptor mutants G185C and R202C by a second site suppressor mutation
The Journal of biological chemistry 2001
read onlineSorting functions of the individual cytoplasmic domains of the G protein-coupled vasopressin V(2) receptor in Madin Darby canine kidney epithelial cells
Molecular pharmacology 2001
read onlineThe role of conserved extracellular cysteine residues in vasopressin V2 receptor function and properties of two naturally occurring mutant receptors with additional extracellular cysteine residues
FEBS letters 2000
read onlineMolecular and conformational features of a transport-relevant domain in the C-terminal tail of the vasopressin V(2) receptor
Molecular pharmacology 2000
read onlineA single negatively charged residue affects the orientation of a membrane protein in the inner membrane of Escherichia coli only when it is located adjacent to a transmembrane domain
The Journal of biological chemistry 1999
read onlinePolarized expression of the vasopressin V2 receptor in Madin-Darby canine kidney cells
Kidney international 1999
read onlineFolding and cell surface expression of the vasopressin V2 receptor: requirement of the intracellular C-terminus
FEBS letters 1998
read onlinePolarized cell surface expression of the green fluorescent protein-tagged vasopressin V2 receptor in Madin Darby canine kidney cells
FEBS letters 1998
read onlineA dileucine sequence and an upstream glutamate residue in the intracellular carboxyl terminus of the vasopressin V2 receptor are essential for cell surface transport in COS.M6 cells
Molecular pharmacology 1998
read onlinePrevention of acute allograft rejection by antibody targeting of TIRC7, a novel T cell membrane protein
Immunity 1998
read onlineMembrane targeting and determination of transmembrane topology of the human vasopressin V2 receptor
The Journal of biological chemistry 1996
read onlineVasopressin V2 receptor mutants that cause X-linked nephrogenic diabetes insipidus: analysis of expression, processing, and function
Molecular pharmacology 1996
read onlineProperties of the human arginine vasopressin V2 receptor after site-directed mutagenesis of its putative palmitoylation site
The Biochemical journal 1996
read onlineIdentification and characterization of two functional domains of the hemolysin translocator protein HlyD
Molecular & general genetics : MGG 1994
read onlineTwo novel mutations in the vasopressin V2 receptor gene in patients with congenital nephrogenic diabetes insipidus
Biochemical and biophysical research communications 1994
read onlineThe Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP) is part of the Forschungsverbund Berlin e.V. (FVB), which legally represents seven non-university research institutes - members of the Leibniz Association - in Berlin.
Leibniz-Forschungsinstitut für Molekulare Pharmakologie im Forschungsverbund Berlin e.V. (FMP)
Campus Berlin-Buch
Robert-Roessle-Str. 10,
13125 Berlin, Germany