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piggyBac Transposon System Modification of Primary Human T Cells
Published on: November 5, 2012
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Precise marker excision system using an animal-derived piggyBac transposon in plants
Ayako Nishizawa-Yokoi1, Masaki Endo, Keishi Osakabe
1Plant Genome Engineering Research Unit, National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki, 305-8602, Japan.
The Plant Journal : for Cell and Molecular Biology
|October 30, 2013
Summary
The piggyBac transposon system effectively excises marker genes in plant cells, enabling precise genome editing. This study demonstrates its high-frequency, footprint-free application in rice, advancing gene targeting technology.
Area of Science:
- Molecular Biology
- Plant Science
- Genetics
Background:
- Gene targeting (GT) in plants requires efficient positive marker excision for desired mutations.
- The piggyBac transposon system is known for successful marker excision in mammals without leaving a footprint.
Purpose of the Study:
- To investigate the functionality of the piggyBac transposon system in plant cells.
- To demonstrate the piggyBac transposon's efficacy in marker excision for plant gene targeting.
Main Methods:
- Developed a transposition assay system using emerald luciferase (Eluc) luminescence in rice cells.
- Utilized hyperactive piggyBac transposase expression in rice calli.
- Confirmed transposition via Polymerase Chain Reaction (PCR), Southern blot, and sequencing.
Main Results:
- Observed Eluc luminescence indicating piggyBac excision in rice calli.
- Demonstrated efficient and precise piggyBac transposition in rice.
- Achieved high-frequency (44.0%) marker gene excision from a reporter locus in T0 plants without re-integration or negative selection.
Conclusions:
- The piggyBac transposon system functions effectively in plant cells for marker excision.
- This system offers a precise and efficient method for footprint-free gene targeting in plants.
- The high excision frequency makes piggyBac a valuable tool for plant genome engineering.
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