Active retrotransposition by a synthetic L1 element in mice

Wenfeng An, Jeffrey S. Han, Sarah Wheelan, Edward S. Davis, Candice E. Coombes, Ping Ye, Christina Triplett, Jef D. Boeke

Research output: Contribution to journalArticle

Abstract

Long interspersed element type 1 (L1) retrotransposons are ubiquitous mammalian mobile elements and potential tools for in vivo mutagenesis; however, native L1 elements are relatively inactive in mice when introduced as transgenes. We have previously described a synthetic L1 element, ORFeus, containing two synonymously recoded ORFs relative to mouse L1. It is significantly more active for retrotransposition in cell culture than all native L1 elements tested. To study its activity in vivo, we developed a transgenic mouse model in which ORFeus expression was controlled by a constitutive heterologous promoter, and we established definitive evidence for ORFeus retrotransposition activity both in germ line and somatic tissues. Germ line retrotransposition frequencies resulting in 0.33 insertions per animal are seen among progeny of ORFeus donor element heterozygotes derived from a single founder, representing a >20-fold increase over native L1 elements. We observe somatic transposition events in 100% of the ORFeus donor-containing animals, and an average of 17 different insertions are easily recovered from each animal; modeling suggests that the number of somatic insertions per animal exceeds this number by perhaps several orders of magnitude. Nearly 200 insertions were precisely mapped, and their distribution in the mouse genome appears random relative to transcription units and guanine-cytosine content. The results suggest that ORFeus may be developed into useful tools for in vivo mutagenesis.

Original languageEnglish (US)
Pages (from-to)18662-18667
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume103
Issue number49
DOIs
StatePublished - Dec 5 2006

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Long Interspersed Nucleotide Elements
Germ Cells
Mutagenesis
Retroelements
Base Composition
Heterozygote
Transgenes
Transgenic Mice
Open Reading Frames
Cell Culture Techniques
Genome

Keywords

  • Gene trap
  • Integration site preference
  • LINE-1
  • Retrotransposon
  • Transgenic mouse

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Active retrotransposition by a synthetic L1 element in mice. / An, Wenfeng; Han, Jeffrey S.; Wheelan, Sarah; Davis, Edward S.; Coombes, Candice E.; Ye, Ping; Triplett, Christina; Boeke, Jef D.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 103, No. 49, 05.12.2006, p. 18662-18667.

Research output: Contribution to journalArticle

An, Wenfeng ; Han, Jeffrey S. ; Wheelan, Sarah ; Davis, Edward S. ; Coombes, Candice E. ; Ye, Ping ; Triplett, Christina ; Boeke, Jef D. / Active retrotransposition by a synthetic L1 element in mice. In: Proceedings of the National Academy of Sciences of the United States of America. 2006 ; Vol. 103, No. 49. pp. 18662-18667.
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