US 12,227,795 B2
Molecular programming tools
Jocelyn Yoshiko Kishi, Cambridge, MA (US); Thomas E. Schaus, Cambridge, MA (US); Peng Yin, Cambridge, MA (US); Feng Xuan, Cambridge, MA (US); Nikhil Gopalkrishnan, Cambridge, MA (US); and Sungwook Woo, Cambridge, MA (US)
Assigned to President and Fellows of Harvard College, Cambridge, MA (US)
Filed by President and Fellows of Harvard College, Cambridge, MA (US)
Filed on Feb. 3, 2022, as Appl. No. 17/592,435.
Application 17/592,435 is a division of application No. 15/999,245, granted, now 11,286,517, previously published as PCT/US2017/018086, filed on Feb. 16, 2017.
Claims priority of provisional application 62/432,017, filed on Dec. 9, 2016.
Claims priority of provisional application 62/429,149, filed on Dec. 2, 2016.
Claims priority of provisional application 62/299,206, filed on Feb. 24, 2016.
Claims priority of provisional application 62/296,310, filed on Feb. 17, 2016.
Prior Publication US 2022/0348990 A1, Nov. 3, 2022
Int. Cl. C12Q 1/68 (2018.01); C12N 15/113 (2010.01); C12N 15/64 (2006.01); C12P 19/34 (2006.01); C12Q 1/6806 (2018.01); C12Q 1/6809 (2018.01); C12Q 1/6811 (2018.01); C12Q 1/6825 (2018.01); C12Q 1/6853 (2018.01)
CPC C12Q 1/6811 (2013.01) [C12N 15/113 (2013.01); C12N 15/64 (2013.01); C12P 19/34 (2013.01); C12Q 1/6806 (2013.01); C12Q 1/6809 (2013.01); C12Q 1/6825 (2013.01); C12Q 1/6853 (2013.01); C12N 2310/122 (2013.01); C12N 2310/127 (2013.01); C12N 2330/50 (2013.01)] 16 Claims
 
1. A method of producing a single-stranded nucleic acid, comprising: combining in reaction buffer
(a) a first catalytic molecule comprising (i) an unpaired 3′ toehold domain, (ii) a paired domain comprising a stopper, and (iii) a linkage domain,
(b) a second catalytic molecule comprising (i) an unpaired 3′ toehold domain, (ii) a paired domain comprising a stopper, and (iii) a linkage domain, wherein the unpaired 3′ toehold domain of the second catalytic molecule binds to a sequence in the paired domain of the first catalytic molecule;
(c) a primer that binds to the unpaired 3′ toehold domain of the first catalytic molecule,
(d) a polymerase having strand displacement activity, and
(e) deoxyribonucleotide triphosphates (dNTPs),
thereby forming a reaction mixture; and
incubating the reaction mixture under conditions that result in nucleic acid polymerization, strand displacement and annealing, for a time sufficient to produce a single-stranded nucleic acid record that is longer than the primer.