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  • Review Article
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Engineering chromosomal rearrangements in mice

Nature Reviews Genetics volume 2, pages 780–790 (2001)Cite this article

Key Points

  • Strategies have recently been developed that allow defined chromosomal rearrangements to be introduced into the mouse genome by engineering them in embryonic stem (ES) cells using the Cre/loxP site-specific recombination system. These rearrangements include chromosomal deletions, duplications, inversions and translocations.

  • Using these strategies, mouse models that accurately recapitulate human chromosomal rearrangements that cause disease have been generated. These include chromosomal translocations that are associated with leukaemia and chromosomal deletions, such as those that cause DiGeorge syndrome and Prader–Willi syndrome.

  • Chromosomal-engineering technology has also been used to generate genetic reagents for the functional analysis of the mouse genome. Deletion chromosomes that are marked by, for example, coat-colour markers, have been engineered to provide segmental haploidy in the mouse genome, and are used to detect recessive mutations that are generated in ethylnitrosourea (ENU) mutagenesis screens.

  • Mouse balancer chromosomes have also been developed using Cre/loxP technology, by tagging chromosomal inversions with recessive lethal mutations and coat-colour markers, for use in ENU mutagenesis screens. These chromosomes facilitate the detection of recessive mutations in the absence of genotypic analysis and allow recessive lethal mutations to be maintained.

  • Although the applications of chromosomal engineering technologies are still in their infancy, they are likely to become invaluable for mapping genetic loci, such as quantitative trait loci, in the future. Marked deletions and balancer chromosomes will also continue to gain importance in large-scale, recessive genetic screens in mice and will significantly contribute to the functional annotation of the mouse genome.

Abstract

The combination of gene-targeting techniques in mouse embryonic stem cells and the Cre/loxP site-specific recombination system has resulted in the emergence of chromosomal-engineering technology in mice. This advance has opened up new opportunities for modelling human diseases that are associated with chromosomal rearrangements. It has also led to the generation of visibly marked deletions and balancer chromosomes in mice, which provide essential reagents for maximizing the efficiency of large-scale mutagenesis efforts and which will accelerate the functional annotation of mammalian genomes, including the human genome.

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Figure 1: A general strategy for chromosomal engineering in mice.
Figure 2: Gene targeting in embryonic stem cells.
Figure 3: Engineering a deletion and/or a duplication in embryonic stem cells.
Figure 4: Engineering an inversion in embryonic stem cells.
Figure 5: Nested chromosomal deletions induced with a retroviral vector.
Figure 6: Engineering chromosomal translocations.
Figure 7: A mouse balancer chromosome and its use in ENU mutagenesis screens.

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Acknowledgements

We thank M. Wentland for comments on this manuscript and A. Pao for assistance in preparing the figures. Work in the authors' lab is supported by the National Institutes of Health, the Howard Hughes Medical Institute and the Wellcome Trust.

Author information

Authors and Affiliations

  1. Department of Molecular and Human Genetics, Program in Developmental Biology, Baylor College of Medicine, One Baylor Plaza, Houston, 77030, Texas, USA

    Yuejin Yu

  2. The Sanger Centre, Hinxton, CB10 1SA, Cambridge, UK

    Allan Bradley

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Supplementary information

Related links

Related links

DATABASES

LocusLink 

AF9

AML1

ETO

MLL

 MGI 

albino

Hprt

Mll

pink-eyed dilution

Trp53

Wnt3

 OMIM 

DiGeorge syndrome

Prader–Willi syndrome

Smith–Magenis syndrome

trisomy 21

FURTHER INFORMATION

Cell line Request Form

Chromosome 11 deletion map

Chromosome 11 ENU mutagenesis programme

Genetic and physical maps of the mouse genome

NCBI's mouse genome sequencing page

STS Physical Map of the Mouse

t(8;21)(q22:q22)

t(9;11)(p22;q23)

The Bradley Lab Protocols

The Sanger Centre

Glossary

SEGMENTAL HAPLOIDY

When a diploid organism is haploid for a certain chromosomal region after its deletion or loss.

BALANCER CHROMOSOME

A chromosome with one or more inverted segments that suppress recombination. They are used as genetic tools because they allow lethal mutations to be maintained without selection.

CROSSING OVER

The exchange of genetic material between two homologous chromosomes.

BLASTOCYST

A preimplantation embryo that contains a fluid-filled cavity called a blastocoel.

POSITIVE SELECTION

When a specific chemical is added to a culture medium, the cells that express a positive selectable marker gene, such as the neomycin or puromycin resistance genes, survive and are selected for.

HPRT MINIGENE

(Hypoxanthine phosphoribosyl transferase gene). This is divided into two complementary, but non-functional, fragments: 5′Hprt contains exons 1–2 and 3′Hprt contains the remaining exons, 3–9. Each Hprt fragment is linked to a loxP site, and Cre-mediated recombination unites the 5′ and 3′ cassettes, and restores Hprt activity, which is required for purine biosynthesis and allows desired recombination events to be selected for in HAT (hypoxanthine, aminopterin and thymidine) medium.

ACENTRIC

A chromosome or chromatid without a centromere.

DICENTRIC

A chromatid or a chromosome that has two centromeres.

K14–AGOUTI

A transgene in which the agouti gene is under the control of the keratin 14 promoter. Its expression produces a yellowish coat colour in mice.

TYROSINASE

Tyrosinase is required for melanin biosynthesis, and the expression of its gene leads to pigment production, and is therefore used as a coat-colour marker.

HSVTK

The herpes simplex virus thymidine kinase (HSVtk) is essential for thymidine nucleotide biosynthesis through a salvage pathway and is often used as a negative selectable marker in gene targeting.

NEGATIVE SELECTABLE MARKER

A negative selectable marker gene, such as HSVtk, allows cells that express it to be killed when a specific chemical is added to a culture medium, whereas cells that no longer express the marker gene survive.

HAPLOINSUFFICIENCY

A phenotype that arises in diploid organisms owing to the loss of one functional copy of a gene.

HYBRID ES-CELL LINE

An embryonic stem (ES) cell line isolated from F1 hybrid embryos, such as from crosses between the strains C57BL/6-Tyrc1Brd × 129S7 or 129S1 × CAST/Ei. These lines facilitate simple sequence length polymorphism analysis.

ENU

(N-ethyl-N-nitrosourea). A potent mutagen that primarily generates single-base-pair mutations in mouse spermatogonia germ cells.

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Yu, Y., Bradley, A. Engineering chromosomal rearrangements in mice. Nat Rev Genet 2, 780–790 (2001). https://doi.org/10.1038/35093564

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