Nicotine has been shown to speed attentional reorienting in cued target detection tasks, and work in young adults suggest that individuals carrying the apolipoprotein E (APOE) e4 allele might show greater sensitivity to the cognitive effects of nicotine. The APOE e4 allele is associated with increased risk of Alzheimer’s disease (AD), and increased sensitivity to nicotine might reflect early cholinergic differences that relate to an enhanced risk of AD. The aim of this study was to investigate effects of nicotine and APOE on attentional reorienting in mid-age participants. APOE e4 (e4+) were compared to non-APOE e4 (e4–) carriers, and functional magnetic resonance imaging (fMRI) data acquired. Neural data showed that nicotine effects, and the network involved in reorienting, was consistent with studies in young adults. Nicotine improved attentional reorienting at the trend level. Although there were no behavioural effects of genotype, genotype effects were present neurally: e4+ showed decreased extrastriate activation, and enhanced effects of nicotine on reorienting in right middle frontal regions. Drug by genotype interactions were present in hippocampal and anterior cingulate regions. These results are consistent with differential sensitivity to nicotine according to APOE status, possibly reflecting abnormal cholinergic function and accelerated cognitive ageing in mid-age e4+.