In this letter, we present a new design that uses single-stranded (ss) DNAs containing two stretches of cytosine (C)-rich domains for the controlled assembly of gold nanoparticles (Au NPs). We show that this assembly is driven by the formation of interparticle i-motif (four-stranded C-quadruplex) structures formed between the C-rich domains of the ssDNAs on neighboring Au NPs. The assembly happens only at slightly acidic pH conditions (pHs below the pK(a) of the i-motif). The assembly is reversible and can be switched by changing the solution pH. The assembly and disassembly process is accompanied by distinct color changes that are clearly visible to the naked eye. This development may have applications in the controlled assembly of reversible pH-sensitive nanostructures and/or devices.