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  • Review Article
  • Published:

Technicolour transgenics: imaging tools for functional genomics in the mouse

Nature Reviews Genetics volume 4, pages 613–625 (2003)Cite this article

Key Points

  • The mouse is the premier mammalian model organism and provides an unparalleled platform for modelling mammalian development and disease. Comprehensive functional annotation of the mouse genome relies on an integrative approach, using an array of tools for unravelling the molecular and cellular basis of normal and mutant phenotypes.

  • Optical-imaging technologies provide a way to perform live-cell analyses in an organismal context. Continued improvements in fluorescence-based imaging technologies allow deeper imaging and better spectral separation of fluorescent-protein reporters in living specimens.

  • Genetically-encoded fluorescent proteins that are expressed in normal and mutant mice represent high-resolution high-contrast multicolour vital markers for monitoring gene activity and protein function. They pave the way for the multidimensional multispectral imaging of living specimens and the generation of multidimensional atlases of model organisms.

  • An increasing number of fluorescent-protein reporters are available for use in mice. These fall into four main categories: spectral-variant reporters, subcellularly localized reporters, photo-activatable reporters and reporters that act as biochemical sensors.

  • The development of static in vitro culture systems for mammalian embryos and explanted tissues promotes normal ex vivo growth, which is necessary for vital-imaging experiments.

  • Researchers now possess a powerful toolbox that offers the potential to visualize and quantify biological processes at the cellular and subcellular level, both in vitro and in intact living organisms.

Abstract

Over the past decade, a battery of powerful tools that encompass forward and reverse genetic approaches have been developed to dissect the molecular and cellular processes that regulate development and disease. The advent of genetically-encoded fluorescent proteins that are expressed in wild type and mutant mice, together with advances in imaging technology, make it possible to study these biological processes in many dimensions. Importantly, these technologies allow direct visual access to complex events as they happen in their native environment, which provides greater insights into mammalian biology than ever before.

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Figure 1: Combinatorial fluorescent-protein-reporter detection in live chimaeras and double transgenics.
Figure 2: An X-linked fluorescent reporter.
Figure 3: Green fluorescence as a marker of branching morphogenesis in the developing kidney.
Figure 4: Fluorescent fusion proteins as markers of subcellular compartments.
Figure 5: Towards a high-resolution multidimensional atlas of a living mouse.

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Acknowledgements

We thank F. Costantini, L. Jones and J. Lichtman for generously providing figures. We apologize to the authors of many important findings and strains of mice that are not discussed here owing to space limitations. Our work is supported by grants from the National Institutes of Health, National Science Foundation and Muscular Dystrophy Association. A.-K.H. is a fellow of the American Heart Association.

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Authors and Affiliations

  1. Department of Genetics and Development, College of Physicians and Surgeons of Columbia University, New York, 10032, USA

    Anna-Katerina Hadjantonakis & Virginia E. Papaioannou

  2. Beckman Institute, California Institute of Technology, Pasadena, 91125, California, USA

    Mary E. Dickinson & Scott E. Fraser

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  1. Anna-Katerina Hadjantonakis
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  2. Mary E. Dickinson
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Correspondence to Anna-Katerina Hadjantonakis.

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Glossary

THERMOLABILE

Unstable at moderate or increased temperatures.

CHIMAERA

An animal that is generated from, and comprises, several genetically distinct populations of cells that are derived from more than one individual.

EMBRYONIC-STEM-CELL-MEDIATED TRANSGENESIS

A method in which DNA is introduced into embryonic stem (ES) cells and integrates randomly, or through gene targeting, into the genome. Transgenic ES cells are delivered to the germline through the generation of (ES cell↔embryo) chimaeras.

FATE MAP

A spatial map of the fates of different embryonic cells at a particular stage of development.

CHORDONEURAL HINGE

A region at the posterior end of a vertebrate embryo that gives rise to both neural and mesodermal cells.

TROPHOBLAST

An extraembryonic lineage that is derived from the trophectoderm of the blastocyst, which gives rise to the fetal portion of the placenta.

PLURIPOTENT

Able to give rise to a wide range of, but not all, cell lineages (usually all fetal lineages and a subset of extraembryonic lineages).

HAEMATOPOIETIC

Giving rise to the cellular elements of the blood, such as the white blood cells, red blood cells and platelets.

MOSAIC

An organism that consists of cells of more than one genotype. The strict definition requires that the genotypically different cells all derive from a single zygote. The term mosaic is also used more broadly to describe any organism comprised of cells of different genotypes.

COELENTERATE

Radially symmetrical invertebrates, which include corals, sea anemones and jellyfish.

FLUOROPHORE

The core portion of a molecule that is directly responsible for absorbing photons.

THERMOSTABLE

Able to withstand moderate heat without the loss of characteristic properties, such as fluorescence.

QUANTUM YIELD

The probability of luminescence occurring under given conditions, which is expressed as the ratio of the number of photons emitted to the number absorbed.

EMBRYONIC STEM CELLS

(ES cells). Stem cells have the dual capacity to self-replicate and differentiate into several specialized derivatives. ES cells are pluripotent cells that are derived from pre-implantation stage (usually blastocyst) mammalian embryos. Mouse ES cells can be propagated and manipulated in vitro, yet still retain their pluripotency.

HEMIZYGOTE

An animal with a transgene insertion on one chromosome of a homologous pair, rather than on each of the two homologous chromosomes (homozygote).

PERDURANCE

The ongoing stability and activity of a protein in the cellular environment.

EPIBLAST

An embryonic lineage that is derived from the inner-cell mass of the blastocyst, which gives rise to the body of the fetus.

PHOTOBLEACHING

The irreversible destruction of a fluorophore that is under illumination.

MUSHROOM BODIES

The region of the Drosophila brain that is required for olfactory learning and memory.

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Hadjantonakis, AK., Dickinson, M., Fraser, S. et al. Technicolour transgenics: imaging tools for functional genomics in the mouse. Nat Rev Genet 4, 613–625 (2003). https://doi.org/10.1038/nrg1126

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