Site-specific DNA excision in transgenic rice with a cell-permeable cre recombinase.
The removal of selected marker genes from transgenic plants is necessary to address biosafety concerns and to carry out further experiments with transgenic organisms. In the present study, the 12-amino-acid membrane translocation sequence (MTS) from the Kaposi fibroblast growth factor (FGF)-4 was used as a carrier to deliver enzymatically active Cre proteins into living plant cells, and to produce a site-specific DNA excision in transgenic rice plants. The process, which made cells permeable to Cre recombinase-mediated DNA recombination, circumvented the need to express Cre under spatiotemporal control and was proved to be a simple and efficient system to achieve marker-free transgenic plants. The ultimate aim of the present study is to develop commercial rice cultivars free from selected marker genes to hasten public acceptance of transgenic crops.
Cao
Ming-Xia
MX
National Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, People's Republic of China.
Huang
Jian-Qiu
JQ
Yao
Quan-Hong
QH
Liu
Sheng-Jun
SJ
Wang
Cheng-Long
CL
Wei
Zhi-Ming
ZM
D015139:Blotting, Southern; D002478:Cells, Cultured; D002934:Cinnamates / Q000494:pharmacology; D018744:DNA, Plant / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D051519:Fibroblast Growth Factor 4 / Q000235:genetics; D017353:Gene Deletion; D015870:Gene Expression / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D006921:Hygromycin B / Q000031:analogs & derivatives / Q000494:pharmacology; D019426:Integrases / Q000235:genetics* / Q000378:metabolism; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D019913:Nuclear Localization Signals / Q000235:genetics; D012275:Oryza / Q000166:cytology / Q000235:genetics*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics / Q000378:metabolism; D018515:Plant Leaves / Q000235:genetics; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D021382:Protein Sorting Signals / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D011995:Recombination, Genetic / Q000187:drug effects / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism
D002934:Cinnamates; D018744:DNA, Plant; D051519:Fibroblast Growth Factor 4; D019913:Nuclear Localization Signals; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D006921:Hygromycin B; C026273:hygromycin A; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase; C045073:Cre recombinase; D019426:Integrases; D005966:Glucuronidase