Now Kosuri\u2019s lab has improved the technique with dye-cycling ORBIT<\/a>, allowing molecular motion to be observed over much longer periods of time (Cell Rep. Methods 2026, DOI: 10.1016\/j.crmeth.2026.101550).<\/p>\n Like the original ORBIT, dye-cycling ORBIT is based on DNA origami<\/a>, a technique where researchers engineer DNA sequences that fold in on themselves to create complex structures. In this case, the DNA structure is a four-armed rotor connected to a shaft that can be ligated to an enzyme of choice. \u201cThe rotor itself consists of about 200 separate strands of DNA,\u201d Kosuri says. \u201cWe just order them, and then we mix them together in right proportions, heat them up, cool them down. And over a couple of days, we have trillions of rotors that we can use.\u201d<\/p>\n The improvement to ORBIT is really simple, Kosuri says. Rather than fixing the fluorescent dyes covalently to the rotor arms, the new rotors have \u201clanding pads\u201d of single-stranded DNA where dyes, tagged with a complementary strand, can transiently bind.<\/p>\n With multiple binding sites on each arm, oligo-dye molecules can continually join and leave the rotor arms. This allows the researchers to track the new dye-cycling ORBIT rotors for much longer compared with the original ORBIT rotors. In one experiment, Kosuri\u2019s team could track transcription for 10 min.<\/p>\n \u201cI dream of doing it for hours on end to see if [enzymes] go through moods and personality changes over time, or if they kind of stay the same,\u201d Kosuri says.<\/p>\n And Kosuri hopes that other researchers will apply this technique to visualize the movement of the proteins they\u2019re most interested in. \u201cI think a lot of the answers to how biology works lies in mechanical motions, not just in the chemical reactions,\u201d he says.<\/p>\n
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