To study protein function, e.g. in the context of genetic diseases, viral vectors are increasingly becoming the method of choice to mediate gene transfer into mammalian cells in vitro and in vivo. For molecular biological applications wild type viral sequences are replaced by foreign DNA and packaged into viral particles. Viral coat proteins determine the serotype as well as the specificity of the gene shuttle. To ensure a high level of biosafety, the replication machinery has been removed from the virus genome and is provided in trans during production only. The resulting viral particles are able to transduce cells and mediate transgene expression, but may not replicate any more.
Today, recombinant viral vectors are used to analyze gene function by either mediating a "knock-in" or rescue a "knock-out" phenotype by translating the gene of interest or by silencing an endogenous gene by inducing RNA interference through transcription of short interfering RNA (shRNA) sequences.
The Vector Core Facility would like to support researchers by selecting the most appropriate viral vector for their experiments. According to their requirements such as the size of the transgene, cell or tissue specificity, different virus types may be preferable. Furthermore, the stability and persistence of transgene expression as well as the virus titers necessary for conducting the project have to be taken into account.
Not every virus is able to infect different cell types with equal efficiency: tropism, infectivity, persistence in the target cell and toxicity sometimes differ significantly. Often is not successful infection in vitro in vivo studies transferable. Using a gene transfer vector, therefore, is highly dependent on the envisaged project objective. If literature searches provide no evidence of the susceptibility of your target cells, as it is in the Vector Facility a selection of virus stocks with reporter genes available for this purpose (see PDF below).
The Vector Facility provides the production of viral gene transfer vectors individually according to customer requirements using appropriate transfer plasmids. The choice of the optimal transfer plasmid is i.a. depending on the size of the transgene, the promoter and the desired duration of expression. In the Vector Facility is a selection of transfer plasmids available that we can offer to insert your gene of interest. When selecting a suitable vehicle, please contact us for discussions. The following recombinant viruses can currently be produced in the Vector Facility:
- Adeno-associated virus: serotype 1, 2, 3, 4, 5, 6, 7, 8, 9, rh10, 12, DJ
- Adenovirus serotype 5
- Integrating and non-integrating y retroviruses: ecotropic, amphotropic, pantropic
- Integrating and non-integrating lentivirus
- Semilki Forest Virus
User Agreement (German only)
Overview promotor collection and AAV tropism
Cetuximab resistance in head and neck cancer is mediated by EGFR-K521 polymorphism. Braig F, Kriegs M, Habel B, Voigtlaender M, Grob T, Biskup K, Blanchard V, Sack M, Thalhammer A, Ben Batalla I, Braren I, Laban S, Danielczyk A, Goletz S, Jakubowicz E, Märkl B, Trepel M, Knecht R, Riecken K, Fehse B, Loges S, Bokemeyer C, Binder M. Cancer Res. 2016 Dec 28. pii: canres.0754.2016. doi: 10.1158/0008-5472.CAN-16-0754. [Epub ahead of print]
Human serum substitution by artificial sera of scalable allergen reactivity based on polyclonal antibodies and chimeras of human FcγRI and IgE domains. Offermann N, Plum M, Hübner U, Rathloff K, Braren I, Fooke M, Spillner E. Allergy. 2016 Sep 2. doi: 10.1111/all.13038. [Epub ahead of print]
Mybpc3 gene therapy for neonatal cardiomyopathy enables long-term disease prevention in mice. Mearini G, Stimpel D, Geertz B, Weinberger F, Krämer E, Schlossarek S, Mourot-Filiatre J, Stoehr A, Dutsch A, Wijnker PJ, Braren I, Katus HA, Müller OJ, Voit T, Eschenhagen T, Carrier L. Nat Commun. 2014 Dec 2;5:5515. doi: 10.1038/ncomms6515 .
Myelin basic protein cleaves cell adhesion molecule L1 and promotes neuritogenesis and cell survival. Lutz D, Loers G, Kleene R, Oezen I, Kataria H, Katagihallimath N, Braren I, Harauz G, Schachner M. J Biol Chem. 2014 May 9;289(19):13503-18. doi: 10.1074/jbc.M113.530238 . Epub 2014 Mar 26.
Repair of Mybpc3 mRNA by 5'-trans-splicing in a Mouse Model of Hypertrophic Cardiomyopathy. Mearini G, Stimpel D, Krämer E, Geertz B, Braren I, Gedicke-Hornung C, Précigout G, Müller OJ, Katus HA, Eschenhagen T, Voit T, Garcia L, Lorain S, Carrier L. Mol Ther Nucleic Acids. 2013 Jul 2;2:e102. doi: 10.1038/mtna.2013.31 .
Rescue of cardiomyopathy through U7snRNA-mediated exon skipping in Mybpc3-targeted knock-in mice. Gedicke-Hornung C, Behrens-Gawlik V, Reischmann S, Geertz B, Stimpel D, Weinberger F, Schlossarek S, Précigout G, Braren I, Eschenhagen T, Mearini G, Lorain S, Voit T, Dreyfus PA, Garcia L, Carrier L. EMBO Mol Med. 2013 Jul;5(7):1060-77. doi: 10.1002/emmm.201202168 . Epub 2013 May 29.
Impact of ANKRD1 mutations associated with hypertrophic cardiomyopathy on contraction parameters of engineered heart tissue. Crocini C, Arimura T, Reischmann S, Eder A, Braren I, Hansen A, Eschenhagen T, Kimura A, Carrier L. Basic Res Cardiol. 2013 May;108(3):349. doi: 10.1007/s00395-013-0349-x . Epub 2013 Apr 10.
Publications acknowledging the vector facility:
p38MAPK/MK2-dependent phosphorylation controls cytotoxic RIPK1 signalling in inflammation and infection. Menon MB, Gropengießer J, Fischer J, Novikova L, Deuretzbacher A, Lafera J, Schimmeck H, Czymmeck N, Ronkina N, Kotlyarov A, Aepfelbacher M, Gaestel M, Ruckdeschel K. Nat Cell Biol. 2017 Oct;19(10):1248-1259. doi: 10.1038/ncb3614. Epub 2017 Sep 18.
TNFα-induced DLK activation contributes to apoptosis in the beta-cell line HIT. Börchers S, Babaei R, Klimpel C, Duque Escobar J, Schröder S, Blume R, Malik MNH, Oetjen E. Naunyn Schmiedebergs Arch Pharmacol. 2017 May 27. doi: 10.1007/s00210-017-1385-0. [Epub ahead of print]
Viral Vector-Based Transduction of Slice Cultures. Wiegert JS, Gee CE, Oertner TG. Cold Spring Harb Protoc. 2017 Feb 1;2017(2):pdb.prot094896. doi: 10.1101/pdb.prot094896
Layer-specific optogenetic activation of pyramidal neurons causes beta-gamma entrainment of neonatal networks. Bitzenhofer SH, Ahlbeck J, Wolff A, Wiegert JS, Gee CE, Oertner TG, Hanganu-Opatz IL. Nat Commun. 2017 Feb 20;8:14563. doi: 10.1038/ncomms14563.
Active cortical dendrites modulate perception. Takahashi N, Oertner TG, Hegemann P, Larkum ME. Science. 2016 Dec 23;354(6319):1587-1590. doi: 10.1126/science.aah6066.
Cardiac repair in guinea pigs with human engineered heart tissue from induced pluripotent stem cells. Weinberger F, Breckwoldt K, Pecha S, Kelly A, Geertz B, Starbatty J, Yorgan T, Cheng KH, Lessmann K, Stolen T, Scherrer-Crosbie M, Smith G, Reichenspurner H, Hansen A, Eschenhagen T. Sci Transl Med. 2016 Nov 2;8(363):363ra148.
An Efficient and Versatile System for Visualization and Genetic Modification of Dopaminergic Neurons in Transgenic Mice. Tillack K, Aboutalebi H, Kramer ER. PLoS One. 2015 Aug doi: 20;10(8):e0136203. 10.1371/journal.pone.0136203 . eCollection 2015.
An improvedchloride-conductingchannelrhodopsin for light-inducedinhibition of neuronalactivity in vivo. Wietek J, Beltramo R, Scanziani M, Hegemann P, Oertner TG, Simon Wiegert J. Sci Rep. 2015 Oct 7;5:14807. doi: 10.1038/srep14807 .
Radixin regulates synaptic GABAA receptor density and is essential for reversal learning and short-term memory. HausratTJ, Muhia M, Gerrow K, Thomas P, Hirdes W, Tsukita S, Heisler FF, Herich L, Dubroqua S, Breiden P, Feldon J, Schwarz JR, Yee BK, Smart TG, Triller A, Kneussel M. Nat Commun. 2015 Apr 20;6:6872. doi: 10.1038/ncomms7872 .
The Vector Facility is open to all researchers / -Innen at the UKE and interested welcome to use. Please contact for further information by email to the contact address (firstname.lastname@example.org) and note the agreement of the Facility.
- Vector Facility