title: Active retrotransposition by a synthetic L1 element in mice.
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.
authors.affiliation: High Throughput Biology Center and Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, 733 North Broadway, Baltimore, MD 21205, USA.
authors.forename: Wenfeng
authors.initials: W
authors.lastname: An
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Han
authors.forename: Sarah J
authors.initials: SJ
authors.lastname: Wheelan
authors.forename: Edward S
authors.initials: ES
authors.lastname: Davis
authors.forename: Candice E
authors.initials: CE
authors.lastname: Coombes
authors.forename: Ping
authors.initials: P
authors.lastname: Ye
authors.forename: Christina
authors.initials: C
authors.lastname: Triplett
authors.forename: Jef D
authors.initials: JD
authors.lastname: Boeke
mesh_terms: D000818:Animals; D005260:Female; D006367:HeLa Cells; D006801:Humans; D020084:Long Interspersed Nucleotide Elements / Q000235:genetics / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D011995:Recombination, Genetic / Q000502:physiology*