Hyperactive mutants of Himar1 transposase and methods for using the same

US6368830B1 · kind B1 · utility

Assignees

• President and Fellows of Harvard College · US
• The Board of Trustees of the University of Illinois · US

Inventors

• David J. Lampe · Glenshaw, US
• Hugh M. Robertson · Urbana, US
• Eric Rubin · Newton, US
• Brian Akerley · Brookline, US

Key dates

• Filing date: Sep 27, 2000
• Grant date: Apr 9, 2002
• Expiry date: Sep 27, 2020

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N15/74
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Mariner-family transposable elements are active in a wide variety of organisms and are becoming increasingly important genetic tools in species lacking sophisticated genetics. The Himar1 element, a member of the mariner family, isolated from the horn fly, Haematobia irritans, is active in Escherichia coli when expressed appropriately. Using this fact, a genetic screen was devised to isolate hyperactive mutants of Himar1 transposase that enhance overall transposition from 4 to 50-fold as measured in an E. coli assay. These hyperactive Himar1 mutant transposases should enable sophisticated analysis of the biochemistry of mariner transposition and should improve efficiency of a variety of genetic manipulations involving transposition in vivo and in vitro such as random mutagenesis or transgenesis in a wide range of host cells than the transposable elements previously available.