Abstract Wavelength‐selective photo‐regulation by multiple chromophores responding to different wavelengths can expand the variation of photo‐manipulating systems. Herein, we report the orthogonal photo‐regulation of duplex formation between serinol nucleic acid (SNA) and RNA using light‐induced crosslinking reactions mediated by a new photo‐reactive nucleobase 8‐naphthylvinyladenine ( NV A) and previously described 8‐pyrenylvinyladenine ( PV A). An intrastrand crosslink was induced in an SNA strand containing two adjacent NV A residues by irradiation with 340–405 nm light; the crosslink was reversed by irradiation with ≤300 nm light. In an SNA strand with adjacent NV A and PV A residues, an intrastrand crosslink resulted from irradiation with 405–465 nm light that was reversed by irradiation with ≤340 nm light. Intrastrand photo‐crosslinking caused severe destabilization of an SNA/RNA duplex, resulting in dissociation to single strands. Cycloreversion resulted in duplex formation. With these NV A/ NV A and NV A/ PV A photo‐switches, four hybridization states of two SNA/RNA duplexes could be orthogonally photo‐controlled by irradiation with a suitable wavelength of light.