organisation - the Instituto Gulbenkian de Ciência

TRANSGENICS
UNIT
Moisés Mallo Head
PhD in Molecular Biology, University of Santiago de Compostela, Spain, 1991
PI of the Patterning and Morphogenesis Group
Head of Facility since 2001
The Transgenics Unit is the gene manipulation unit. The unit provides transgenic
services to research staff. Both classic transgenics, by pronuclear microinjection,
and gene targeting, by blastocyst injection of embryonic ES cells, are
offered as a service. The unit also offers the cryopreservation of embryos for
long-term storage.
FACILITY STAFF
Ana Nóvoa (Research Technician)
MAJOR PROJECTS AND ACCOMPLISHMENTS
The work of the unit during 2011 mostly involved the genesis of transgenic mice
by pronuclear DNA injection. Most transgenics were produced by microinjection
into the FVB/N background. This year, we produced transgenic lines from
four different constructs. One of them was a BAC reporter. The other three
generated tools for conditional inactivation studies in mesodermal derivatives
and for labelling embryonic muscle precursors with fluorescent proteins to allow
their isolation from the living embryo using cell sorting approaches. The
rest of the transgenic production was not aimed at generating transgenic lines
because transgene expression resulted in embryonic lethal phenotypes. Therefore,
these injections were designed for studies using a transient transgenic
approach. During this year, we created transgenic embryos from more than 20
different constructs, including two different BACs. Similar to previous years, the
transgenic efficiency was kept high, being around 30% for normal transgenics
and 17% for transgenics injecting BAC constructs. The transgenic unit also produced
four chimeric mice from one strain of ES-cells by injection into C57BL/6
blastocysts. All these animals contained higher than 70% chimerism.
EQUIPMENT AND INFRASTRUCTURE
• 1 Microinjection setup with Nikon inverted microscope equipped with DIC
optics, and three dimensional Narishige micromanipulators;
• 1 Microinjection setup with Leica inverted microscope equipped with DIC
optics, and three dimensional, power assisted, Narishige micromanipulators;
• 2 FemtoJet pump;
• 1 Sutter P-87 Flaming/Brown micropipette puller;
• 1 Zeiss SV6 Stereomicroscope with training head;
• 2 Standard Zeiss SV6 Stereomicroscopes;
• 1 CO2 incubator;
• 1 Ultrasonic Cleaning Device.
Funding Calouste Gulbenkian Foundation and Fundação para a Ciência e a
Tecnologia, Portugal
TwoCellEmbryos - Mouse embryos at the two-cell stage.
IGC ANNUAL REPORT ‘11
FACILITIES AND SERVICES
87