US 12,291,742 B2
Selective oxidation of 5-methylcytosine by TET-family proteins
Anjana Rao, La Jolla, CA (US); Mamta Tahiliani, New York, NY (US); Kian Peng Koh, Jamaica Plain, MA (US); Suneet Agarwal, Belmont, MA (US); and Aravind Iyer, Bethesda, MD (US)
Assigned to The Children's Medical Center Corporation, Boston, MA (US); and The United States of America, As Represented by the Secretary, Department of Health & Human Services, Bethesda, MD (US)
Filed by The Children's Medical Center Corporation, Boston, MA (US); and The United States of America, As Represented by the Secretary, Department of Health & Human Services, Bethesda, MD (US)
Filed on Feb. 27, 2024, as Appl. No. 18/588,987.
Application 18/588,987 is a continuation of application No. 17/675,502, filed on Feb. 18, 2022, granted, now 12,018,320.
Application 17/675,502 is a continuation of application No. 17/350,181, filed on Jun. 17, 2021, abandoned.
Application 17/350,181 is a continuation of application No. 16/380,846, filed on Apr. 10, 2019, granted, now 11,072,818, issued on Jul. 27, 2021.
Application 16/380,846 is a continuation of application No. 15/440,815, filed on Feb. 23, 2017, granted, now 10,323,269, issued on Jun. 18, 2019.
Application 15/440,815 is a continuation of application No. 15/341,344, filed on Nov. 2, 2016, granted, now 10,533,213, issued on Jan. 14, 2020.
Application 15/341,344 is a continuation of application No. 15/193,796, filed on Jun. 27, 2016, granted, now 10,443,091, issued on Oct. 15, 2019.
Application 15/193,796 is a continuation of application No. 13/795,739, filed on Mar. 12, 2013, granted, now 9,447,452, issued on Sep. 20, 2016.
Application 13/795,739 is a continuation of application No. 13/120,861, granted, now 9,115,386, issued on Aug. 25, 2015, previously published as PCT/US2009/058562, filed on Sep. 28, 2009.
Claims priority of provisional application 61/121,844, filed on Dec. 11, 2008.
Claims priority of provisional application 61/100,995, filed on Sep. 29, 2008.
Claims priority of provisional application 61/100,503, filed on Sep. 26, 2008.
Prior Publication US 2024/0254542 A1, Aug. 1, 2024
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/26 (2006.01); C12N 9/10 (2006.01); C12Q 1/6806 (2018.01); C12Q 1/6827 (2018.01); C12Q 1/6869 (2018.01); G01N 33/53 (2006.01)
CPC C12Q 1/6827 (2013.01) [C12N 9/1007 (2013.01); C12Q 1/26 (2013.01); C12Q 1/6806 (2013.01); C12Q 1/6869 (2013.01); G01N 33/5308 (2013.01); C12N 2501/70 (2013.01); C12N 2501/71 (2013.01); C12N 2501/724 (2013.01); C12Q 2521/531 (2013.01); C12Q 2522/10 (2013.01); C12Q 2537/164 (2013.01); C12Q 2600/154 (2013.01)] 30 Claims
OG exemplary drawing
 
1. A method comprising:
(a) contacting a nucleic acid molecule comprising cytosine and 5-methylcytosine with an isolated enzyme comprising a TET polypeptide, a catalytically active fragment thereof, a variant thereof, or any combination thereof in an amount effective to convert 5-methylcytosine of said nucleic acid molecule to an oxidized derivative thereof;
(b) converting cytosine of said nucleic acid molecule to uracil; and
(c) conducting a sequencing reaction to obtain a sequence of said nucleic acid molecule.