Here, we report one-step DNA functionalization of hydrophobic iron oxide nanoparticles (IONPs) using DNA-grafted poly(acrylic acid) (PAA-g-DNA). PAA-g-DNA was synthesized by coupling PAA to amine-modified oligonucleotides via solid-phase amide chemistry, which yielded PAA grafted with multiple DNA strands with high graft efficiencies. Synthesized PAA-g-DNA was utilized as a phase-transfer and DNA functionalization agent for hydrophobic IONPs, taking advantage of unreacted carboxylic acid groups. The resulting DNA-modified IONPs were well dispersed in aqueous solutions and possessed DNA binding properties characteristic of polyvalent DNA nanostructures, showing that this approach provides a simple one-step method for DNA functionalization of hydrophobic IONPs.