These include the AP2 ERF transcription factors DREB2A, DREB2B, and CBF4, AtMYB2, which functions in concert with AtMYC2, and the NAC family transcription factors ANAC19, Enzastaurin Phase 3 ANAC055 ATNAC3, and ANAC072 RD26, which at least par tially function in concert with a zinc finger homeodo main protein Inhibitors,Modulators,Libraries ZFHD1 . Therefore, while overexpression of ABF3 affects one of the key drought response pathways, it is not the only pathway mediating the drought response at the gene expression level. The use of the Cre lox system to create control lines also created an opportunity to examine the effects of the Cre lox system on the transcriptome. Site specific recombination technologies can be used to excise select able markers or other undesirable genetic elements and can also be used to direct site specific integration of transgenes.
While many studies have employed Cre mediated recombination in plant systems with no apparent unintended effects, other studies have observed a range of abnormal phenotypes including Inhibitors,Modulators,Libraries growth defects, leaf chlorosis, delayed flowering, and male sterility. In tobacco plants transformed with a chloroplast targeted Cre recombinase, recombina tion was observed involving Inhibitors,Modulators,Libraries cryptic lox sites in the plas tid genome, but invariably the second lox site was located within the transgene. These recombina tions could result in deletions of up to 147 kb, but they did not cause any deleterious effects in the plants. Studies in animal systems have similarly revealed that Cre recombinase can have unintended effects, often leading to chromosomal aberrations.
These studies suggest that the Cre lox system has the potential to cause unintended effects in plant systems, by mediating recombination with cryptic lox sites that may be present in the genome resulting in large dele tions. Inhibitors,Modulators,Libraries Such cryptic lox sites are difficult to identify since they may deviate substantially from conventional loxP sites and not all of the unintended Inhibitors,Modulators,Libraries effects may pro duce readily apparent phenotypic abnormalities, so studying the unintended effects of Cre recombinase using a non targeted approach such as microarray analy sis is essential for establishing the utility and safety of this technology. In this study, we performed microarray analysis on Arabidopsis plants engineered to be drought tolerant through overexpression of the transcription factor ABF3 with the goal of identifying unintended pleiotropic effects. The results suggest that overexpression of ABF3 has a minimal impact on the transcriptome, with differ ences in the gene expression pattern only detectable in response to drought and then being suggestive of tran scriptional reprogramming selleck chemicals llc as opposed to the activation of novel pathways.
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