Methods and compositions for transposition using minimal segments of the eukaryotic transformation vector piggyBac

US10087463B2 · kind B2 · utility

Assignee

• University of Notre Dame du Lac · US

Inventors

• Malcolm J. Fraser · Granger, US
• Xu Li · 林湖乡, CN

Key dates

• Filing date: Jun 19, 2006
• Grant date: Oct 2, 2018
• Expiry date: Jun 27, 2027

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N2800/90
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The present invention provides a method for transforming an insect genome that has a much enhanced transformation frequency. The vectors and plasmids employed in the method are further described as transposition vectors that include a minimal amount of nucleotide sequence homologous to a 5′ region and a 3′ region of a native piggyBac nucleic acid sequence. The transformed cells or embryos may also be developed into transgenic organisms. Disclosed are minimal piggyBac-based plasmid constructs that comprises a minimal nucleic acid sequence homologous to a 5′ end of a piggyBac nucleic acid sequence (about 60-80 bp, particularly 66 bp) and a relatively long (300 to about 380 bp, particularly 311 bp or 378 bp) continuous nucleic acid sequence homologous to a 3′ end of a piggyBac native nucleic acid sequence. Methods employing these constructs include the use of a helper plasmid. Transformation frequencies employing the constructs are enhanced 100-fold or higher over that transformation frequency obtained using other than the herein described constructs.