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 drivenby the formation of interparticle i-motif (four-stranded C-quadruplex) structures formed between the C-rich domainsof the ssDNAs on neighboring Au NPs. The assembly happens only at slightly acidic pH conditions (pHs below thepKa of the i-motif). The assembly is reversible and can be switched by changing the solution pH. The assembly anddisassembly process is accompanied by distinct color changes that are clearly visible to the naked eye. This developmentmay have applications in the controlled assembly of reversible pH-sensitive nanostructures and/or devices.