Simplifying transgene locus structure through Cre-lox recombination.
Transgene silencing is often associated with multicopy integrations, which occur frequently during plant transformation. Transgene expression can be restored in a number of multicopy loci by converting them to single copy. This chapter describes a plant transformation protocol based on use of the Cre-lox system, which allows conversion of a multicopy transgene locus into single copy. The strategy is based on designing a transformation vector with lox sites, developing transgenic lines, and introducing Cre activity to initiate Cre-lox recombination, which leads to the simplification of a multicopy locus to a single- or low-copy state. This method is compatible with both gene gun and Agrobacterium-mediated gene delivery and should be particularly useful for crops that are difficult to transform.
Department of Crop, Soil and Environmental Sciences, University of Arkansas, Fayetteville, AR, USA, vibhas@uark.edu.
Vibha
V
Srivastava
David W
DW
Ow
D060054:Agrobacterium / Q000235:genetics*; D018556:Crops, Agricultural / Q000235:genetics; D018014:Gene Transfer Techniques; D056426:Genetic Loci; D005822:Genetic Vectors / Q000235:genetics; D019426:Integrases / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics; D019076:Transgenes
C045073:Cre recombinase; D019426:Integrases