US 11,718,843 B2
Light-inducible targeted modification of nucleic acids and genetic information
Stefan Schillberg, Aachen (DE); Gerda Noelke, Aachen (DE); and Stefano Di Fiore, Neuss (DE)
Assigned to Fraunhofer-Gesellschaft zur Foerderung der angewandten Forschung e.V., Munich (DE)
Appl. No. 16/772,028
Filed by Fraunhofer-Gesellschaft zur Foerderung der angewandten Forschung e.V., Munich (DE)
PCT Filed Dec. 19, 2018, PCT No. PCT/EP2018/085860
§ 371(c)(1), (2) Date Jun. 11, 2020,
PCT Pub. No. WO2019/121928, PCT Pub. Date Jun. 27, 2019.
Claims priority of application No. 17208812 (EP), filed on Dec. 20, 2017.
Prior Publication US 2021/0214710 A1, Jul. 15, 2021
Int. Cl. C12N 15/01 (2006.01); C12N 13/00 (2006.01); G01N 21/64 (2006.01); C12N 15/10 (2006.01)
CPC C12N 13/00 (2013.01) [C12N 15/01 (2013.01); C12N 15/102 (2013.01); C12Q 2523/313 (2013.01)] 5 Claims
 
1. A method of breaking a target nucleic acid in vitro, the method comprising:
(i) providing, in vitro, a target nucleic acid sequence comprising plasmid or linear DNA to be broken;
(ii) providing a pre-assembled complex for breaking the target nucleic acid sequence, the complex comprising:
a. a metal nanocluster of 2 to 14 silver atoms;
b. a nucleic acid molecule of single stranded DNA that is different from the target nucleic acid sequence and shorter than the target nucleic acid sequence, the single stranded DNA comprising:
i. an annealing region of 10 to 25 consecutive nucleotides that comprises 60% adenine and thymine nucleotides that are complementary to consecutive nucleotides of the target nucleic acid sequence, and
ii. a nucleation region that flanks the annealing region and is positioned around the metal nanocluster, the nucleation region comprising 8 consecutive cytosine nucleotides;
(iii) contacting the consecutive nucleotides of the annealing region of the complex to the consecutive nucleotides of the target nucleic acid, thereby annealing the pre-assembled complex to the target nucleic acid sequence; and
(iv) inducing a nucleic acid break in the target nucleic acid by exposing the metal nanocluster at the nucleation region to electromagnetic radiation.