# US 11,999,995 B2 · Methylome profiling in animals and uses thereof

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- Number: US 11,999,995 B2
- Document ID: US11999995B2
- Status: Granted
- Kind: B2
- Application number: 16/638,454
- Filed: 2018-08-31
- Published: 2020-11-19
- Granted: 2024-06-04
- Earliest claimed: 2017-08-31
- Assignees: The Regents of the University of California
- Applicants: The Regents of the University of California
- Inventors: Tina Wang; Trey Ideker
- Web page: https://patentgrep.com/patent/US11999995B2

## CPC

- C12Q 1/6827: for detection of mutation or polymorphism
- C12Q 1/6855: Ligating adaptors
- C12Q 1/686: Polymerase chain reaction [PCR]
- C12Q 2523/125: Bisulfite(s)
- C12Q 2535/122: Massive parallel sequencing
- C12Q 2537/143: Multiplexing, i.e. use of multiple primers or probes in a single reaction, usually for simultaneously analyse of multiple analysis
- C12Q 2600/154: Methylation markers
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

The disclosure provides for methylome profiling of animals and further using DNA methylation as a quantitative predictor of the age of an animal.

## Claims

### Claim 1

A method of generating a methylome profile of a canine subject, the method comprising:
isolating and fractioning genomic DNA (gDNA) from a canine subject to form gDNA fragments;
treating the gDNA fragments with bisulfite to convert unmethylated cytosines into uracils;
capturing bisulfite treated gDNA fragments having CpG sites with capture probes having a sequence complementary to bisulfite treated gDNA fragments,
wherein the capture probes that target genetic loci having a genomic location selected from the following:
Genomic location
chromosome
start
end
10
32155572
32155573
10
47400603
47400604
11
12143642
12143643
1
113745958
113745959
11
56810628
56810629
11
56810721
56810722
12
63488774
63488775
14
22067955
22067956
14
40131029
40131030
14
40366402
40366403
16
46562076
46562077
16
53697863
53697864
16
53697881
53697882
17
62480024
62480025
21
27195140
27195141
21
27396898
27396899
22
1780265
1780266
23
42484424
42484425
24
36795647
36795648
25
5057162
5057163
25
5057174
5057175
26
5250703
5250704
26
5330973
5330974
26
5330978
5330979
26
5330998
5330999
26
5331204
5331205
26
5331230
5331231
28
38491744
38491745
30
14008101
14008102
3
15906095
15906096
3
15906160
15906161
32
30355909
30355910
33
25412867
25412868
34
7326879
7326880
37
22520990
22520991
38
18152079
18152080
38
18152126
18152127
38
18152133
18152134
5
29282901
29282902
5
32970662
32970663
5
32970672
32970673
5
48883108
48883109
5
48883118
48883119
5
67246847
67246848
5
79537151
79537152
7
80969482
80969483
9
19960393
19960394
9
20510871
20510872
9
24868907
24868908
9
24868926
24868927
9
24869027
24869028
wherein the capture probes further comprise methylated sequencing adaptors, and
wherein the CpG sites of the bisulfite treated gDNA fragments are orthologous between humans and canines;
amplifying and sequencing the capture gDNA fragments to generate sequencing reads of the canine subject; and
generating the methylome profile of the canine subject from the sequencing reads.

### Claim 2 (depends on 1)

The method of claim 1, wherein the capture probes target 1-10,000 orthologous CPG sites between humans and canines.