Evaluation of seven promoters to achieve germline directed Cre-lox recombination in Arabidopsis thaliana.
Site-specific recombination systems, such as Cre-lox from bacteriophage P1, have become very important tools for plant genome engineering. In many cases a constitutive promoter is used to express the recombinase gene. However, for certain research and commercial applications constitutive Cre-mediated recombination may not be desirable. We have evaluated the potential of seven different germline promoter:cre fusions to remove a stably integrated lox cassette through Cre-mediated recombination in Arabidopsis thaliana. We monitored the functionality of each promoter in the germline of primary transformants by analyzing the presence of the recombined lox cassette in T(2) progeny. The selected germline promoters are involved in different developmental cues, including early stem cell identity (CLAVATA3), flower meristem identity (LEAFY, APETALA1), floral organ identity (AGAMOUS), and meiosis (SOLO DANCERS, DMC1, SWITCH1). For five out of these seven promoters we were able to show that efficient Cre-mediated recombination does, indeed, occur and that the recombination takes place at some point during germline development. Furthermore, a recombination efficiency of 100% is obtained when Cre-expression is regulated by the CLAVATA3 promoter. In addition, with these promoters, we observe much less variation in recombination frequency than previously reported for the 35S promoter. For these reasons, we believe that germline-specific Cre-lox recombination provides an additional tool to the site-specific recombination technology in plants.
Van Ex
Frédéric
F
Laboratory of Plant Genetics, Institute for Molecular Biology and Biotechnology, Vrije Universiteit Brussel (VUB), Pleinlaan 2, 1050 Brussels, Belgium. fvex@vub.ac.be
Verweire
Dimitri
D
Claeys
Martine
M
Depicker
Ann
A
Angenon
Geert
G
D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D005818:Genetic Engineering / Q000379:methods; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic
C045073:Cre recombinase; D019426:Integrases