Discrete,　three-dimensional　(3D)　gold　nanoparticle　(AuNP)　tetrahedron　nanoarchitectures　are　successfully　self-assembled　with　DNA　origami　as　template　with　high　purity　(＞85%).　A　distinct　plasmonic　chiral　response　is　experimentally　observed　from　the　AuNP　tetrahedron　nanoarchitectures　and　appears　in　a　configuration-dependent　manner.　The　chiral　optical　properties　are　then　rationally　engineered　by　modifying　the　structural　parameters　including　the　AuNP　size　and　interparticle　distance.　Theoretical　study　of　the　AuNP　tetrahedron　nanoarchitectures　shows　the　dependence　of　the　chiral　optical　property　on　the　AuNP　size　and　interparticle　distance,　consistent　with　the　ensemble　averaged　measurements.　©　2014　American　Chemical　Society.