Long-range chromosomal engineering is more efficient in vitro than in vitro

Lisa E. Olson; Jason Tien; Sarah South; Roger H. Reeves

doi:10.1007/s11248-005-0389-6

Long-range chromosomal engineering is more efficient in vitro than in vitro

Lisa E. Olson, Jason Tien, Sarah South, Roger H. Reeves

School of Medicine

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

Cre/LoxP mediated chromosomal engineering in embryonic stem (ES) cells has a variety of applications, including the creation of model systems for studying aneuploidy. Targeted meiotic recombination (TAMERE) was proposed as a high efficiency in vivo alternative to effect Cre-mediated recombination, in which Cre recombinase under control of the Synaptonemal Complex 1 promoter is expressed during male meiosis in transgenic mice. TAMERE has been successfully used with LoxP sites up to 100 kb apart. We tested TAMERE for a chromosome engineering application in which LoxP sequences were integrated into sites 3.9 Mb apart on the same (cis) or opposite (trans) copies of mouse Chromosome 16 (MMU16). TAMERE was ineffective in generating either a deletion or a translocation in vivo. The TAMERE method may be of limited use for large genomic rearrangements. The desired translocation was achieved with an in vitro method that can be used in any ES cell line. Mice produced from the reciprocal duplication/deletion of MMU16 in a region homologous to human chromosome 21 provide models that are useful in studies of Down syndrome.

Original language	English (US)
Pages (from-to)	325-332
Number of pages	8
Journal	Transgenic Research
Volume	14
Issue number	3
DOIs	https://doi.org/10.1007/s11248-005-0389-6
State	Published - Jun 2005

Keywords

Chromosome engineering
Cre/LoxP
Down syndrome
TAMERE
Targeted meiotic recombination

ASJC Scopus subject areas

Genetics
Animal Science and Zoology
Agronomy and Crop Science
Biotechnology

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10.1007/s11248-005-0389-6

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title = "Long-range chromosomal engineering is more efficient in vitro than in vitro",

abstract = "Cre/LoxP mediated chromosomal engineering in embryonic stem (ES) cells has a variety of applications, including the creation of model systems for studying aneuploidy. Targeted meiotic recombination (TAMERE) was proposed as a high efficiency in vivo alternative to effect Cre-mediated recombination, in which Cre recombinase under control of the Synaptonemal Complex 1 promoter is expressed during male meiosis in transgenic mice. TAMERE has been successfully used with LoxP sites up to 100 kb apart. We tested TAMERE for a chromosome engineering application in which LoxP sequences were integrated into sites 3.9 Mb apart on the same (cis) or opposite (trans) copies of mouse Chromosome 16 (MMU16). TAMERE was ineffective in generating either a deletion or a translocation in vivo. The TAMERE method may be of limited use for large genomic rearrangements. The desired translocation was achieved with an in vitro method that can be used in any ES cell line. Mice produced from the reciprocal duplication/deletion of MMU16 in a region homologous to human chromosome 21 provide models that are useful in studies of Down syndrome.",

keywords = "Chromosome engineering, Cre/LoxP, Down syndrome, TAMERE, Targeted meiotic recombination",

author = "Olson, {Lisa E.} and Jason Tien and Sarah South and Reeves, {Roger H.}",

note = "Funding Information: The authors thank Dr Wendy Kimber and Stephanie Kane for assistance with the SLK2 vector, Anthony Cukras for sequence analysis of the Mx region, and the Johns Hopkins Transgenic Core for ES cells and blastocyst injection. pCAGGS-Cre was kindly provided by R. Kucherlapati and pBS500 by B. Sauer. LEO was supported by a Howard Hughes Predoctoral Fellowship. This work was supported by PHS awards HD24605 and HD38384 (RHR).",

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AU - Tien, Jason

AU - South, Sarah

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Long-range chromosomal engineering is more efficient in vitro than in vitro

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