research facilities
facilities
Core laboratory of Animal engineering
Integration of genes into the mouse genome represents a powerful experimental approach to study normal as well as altered gene
expression.
It also contributes to identify genes essential for normal development and to generate animal models for human diseases.
The injection room is fully equipped to carry out the entire procedure of making transgenic mice by pronuclear microinjection. Direct microinjection of DNA into the male pronucleus of a mouse zygote is the method most extensively used in the production of transgenic mice. Once the foreign DNA integrates into the mouse chromosomal DNA at the one-cell stage, the transgenic animal will contain the injected DNA in every cell, including those of the germ line, thus allowing transmission of the transgene and the generation of a transgenic mouse line.
The Transgenic Mouse Facility also includes several animal rooms, thus providing housing and breeding space for the mice involved in the transgenic projects, both generated in the Department of Pharmacological Sciences and in other Research Centers.
Selected references
- Sacco MG, Zecca L, Bagnasco L, Chiesa G, Parolini C, Bromley P, Mira Cato' E, Roncucci R, Clerici LA, Vezzoni P. A transgenic mouse model for the detection of cellular stress induced by toxic compounds. Nat Biotechnol 1997, 15: 1392-1397.
- Chiesa G, Stoltzfus LJ, Michelagnoli S, Bielicki JK, Santi M, Forte TM, Sirtori CR, Franceschini G, Rubin EM. Elevated triglycerides and low HDL cholesterol in transgenic mice expressing human apolipoprotein A-IMilano.Atherosclerosis 1998, 136: 139-146, 1998
- Chiesa G, Parolini C, Canavesi M, Colombo N, Sirtori CR, Fumagalli R, Franceschini G, Bernini F. Human apolipoproteins A-I and A-II in cell cholesterol efflux: studies with transgenic mice. Arterioscler Thromb Vasc Biol. 1998, 18: 1417-1423.
- Sacco MG, Caniatti M, Mira Cato' E, Frattini A, Chiesa G, Ceruti R, Adorni F, Zecca L, Scanziani E, Vezzoni P. Liposome-delivered angiostatin strongly inhibits tumor growth and metastatization in a transgenic model of spontaneous breast cancer.Cancer Res. 2000, 60: 2660-2665.
- Parolini C, Chiesa G, Zhu Y, Forte T, Caligari S, Gianazza E, Sacco MG, Sirtori CR, Rubin EM. Targeted replacement of mouse apolipoprotein A-I with the human apoA-I or the mutant apoA-IMilano: evidence of apoA-IM impaired hepatic secretion.J Biol Chem. 2003, 278:4740-4746.
- Parolini C, Chiesa G, Gong E, Caligari S, Cortese MM, Koga T, Forte TM, and Rubin EM. Apolipoprotein A-I and the molecular variant apoA-I Milano: evaluation of the antiatherogenic effects in a knock-in mouse model.Atherosclerosis 2005, 183: 222-229.
Transgenic Operative Platform on Animal Engineering
A Transgenic Operative Platform (TOP) on animal engineering is active at the Department of Pharmacological Sciences. TOP research is oriented on the generation of reporter mice for molecular imaging of biological processes. Molecular imaging is a novel discipline bridging two quite well developed fields of study: molecular biology and computer-aided imaging. This new discipline represents a powerful tool that holds the promise to profoundly change our current approaches to biological investigations. Indeed, appropriately designed “reporter probes” allow to see in real time the extent of transcriptional activity of single genes, the dynamics of intracellular interactions among proteins, the activity of a given drug in an entire organism, the state of activation of receptors or enzymes.
The platform:
- perform active research aimed at generating novel reporters suitable for functional imaging to be used in cell and animal engineering
- provide a technological support in molecular imaging and act as a consultant to scientists working in Academia and Industry
- organize courses for permanent education inthe field of molecular biology applied to imaging (supported by two European Network of Excellence on Imaging, EMIL and DIMI)
- organize a bank of reporter genes and vectors appropriately engineered for imaging purposes
TOP integrates its work with Companies developing imaging instruments and imaging chemicals and experts in bioinformatics for the design of novel, suitable imaging softwares.
The platform has access to a Berthold Night Owl CCD-camera for optical imaging and to a magnetic resonance imaging unit (Bruker) to perform experiments on the reporter mice generated.
Selected references
- Ciana P, Raviscioni M, Mussi P, Vegeto E, Que I, Parker MG, Lowik C, Maggi A. In vivo imaging of transcriptionally active estrogen receptors. Nat Med. 2003, 9:82-86.
- Maggi A, Ottobrini L, Biserni A, Lucignani G, Ciana P. Techniques: reporter mice - a new way to look at drug action. Trends Pharmacol Sci. 2004, 25:337-342.
- Maggi A, Ciana P. Reporter mice and drug discovery and development.Nat Rev Drug Discov. 2005, 4:249-255.
- Ciana P, Fumagalli M, Trincavelli ML, Verderio C, Rosa P, Lecca D, Ferrario S, Parravicini C, Capra V, Gelosa P, Guerrini U, Belcredito S, Cimino M, Sironi L, Tremoli E, Rovati GE, Martini C, Abbracchio MP. The orphan receptor GPR17 identified as a new dual uracil nucleotides/cysteinyl-leukotrienes receptor. EMBO J. 2006, 25:4615-4627.