US 12,467,080 B2
PNA oligomer, use thereof for detecting DNA methylation, and method for detecting DNA methylation using same
Bat Ochir Chinbayar, Daejeon (KR); and Hee Kyung Park, Daejeon (KR)
Assigned to SEASUNBIO MATERIALS, Daejeon (KR)
Appl. No. 17/433,248
Filed by SEASUNBIO MATERIAL, Daejeon (KR)
PCT Filed Sep. 1, 2020, PCT No. PCT/KR2020/011692
§ 371(c)(1), (2) Date Aug. 24, 2021,
PCT Pub. No. WO2021/045479, PCT Pub. Date Mar. 11, 2021.
Claims priority of application No. 10-2019-0108251 (KR), filed on Sep. 2, 2019.
Prior Publication US 2022/0162681 A1, May 26, 2022
Int. Cl. C12Q 1/68 (2018.01); C12Q 1/6827 (2018.01); C12Q 1/686 (2018.01); C12Q 1/6876 (2018.01)
CPC C12Q 1/6827 (2013.01) [C12Q 1/686 (2013.01); C12Q 1/6876 (2013.01); C12Q 2600/154 (2013.01)] 9 Claims
 
1. A method for detecting a methylated gene using the PNA probe represented by the following Formula 1, which comprises:
preparing a first mixture comprising a non-methylated gene and the PNA probe represented by the following Formula 1 which is able to specifically bind to a base sequence of the non-methylated gene;
preparing a second mixture comprising a target gene for methylation analysis, which comprises the same base sequence as the base sequence of the non-methylated gene, and the PNA probe represented by the following Formula 1;
subjecting the first mixture and the second mixture to a polymerase chain reaction (PCR); and
measuring a cycle threshold (ΔCt) value of the PCR:
[ΔCt=Ct(Target Gene for Methylation Analysis)−Ct(Non-methylated Gene Comprising the Same Base Sequence as Target Gene for Methylation Analysis)]

OG Complex Work Unit Chemistry
wherein,
R1 is a hydrophobic substituent, R2 is hydrogen or a hydrophobic substituent, R3 is a hydroxyl group or a hydrophobic substituent,
Base is any one base selected from natural or non-natural nucleobases wherein the natural bases comprise adenine, thymine, guanine, cytosine, and uracil, and the non-natural nucleobases comprise purine, 2,6-diaminopurine, 7-deazaadenine, 7-deazaguanine, N4N4-ethanocytosine, N6N6-ethano-2,6-diaminopurine, 5-methylcytosine, 5-(C3C6)-alkanyluracil, 5-(C3C6)-alkynylcytosine, 5-fluorurocil, and pseudoisocytosine, n is an integer ranging from 5 to 30, and respective structural units included in the PNA probe are able to be the same or different from each other.