CLEAVABLE DISULFIDE LINKERS AND USES THEREOF
Disclosed herein, inter alia, are methods and cleavable compounds that minimize byproduct formation following cleavage in situ.
1 . A cell or tissue comprising a compound having the formula:
wherein
B is a divalent nucleobase;
R 1 is a polyphosphate moiety, monophosphate moiety, or nucleic acid moiety;
R 2 is hydrogen or —OH;
R 3 is a reversible terminator moiety, hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;
R 4 is a detectable moiety;
L 100 is a divalent linker comprising
wherein
R 5 and R 6 are independently unsubstituted alkyl, —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SO 3 H, —SO 4 H, —SO 2 NH 2 , —NHNH 2 , —ONH 2 , substituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; and
W 1 and W 2 are —O—.
2 . The compound of claim 1 , wherein R 2 is hydrogen.
3 . The compound of claim 1 , wherein R 1 is a triphosphate moiety.
4 . The compound of claim 1 , wherein B is a cytosine or a derivative thereof, guanine or a derivative thereof, adenine or a derivative thereof, thymine or a derivative thereof, uracil or a derivative thereof, hypoxanthine or a derivative thereof, xanthine or a derivative thereof, 7-methylguanine or a derivative thereof, 5,6-dihydrouracil or a derivative thereof, 5-methylcytosine or a derivative thereof, or 5-hydroxymethylcytosine or a derivative thereof.
5 . The cell or tissue of claim 1 , wherein R 3 is a reversible terminator moiety.
6 . The cell or tissue of claim 1 , wherein R 3 is
wherein R 7 is substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl.
7 . The cell or tissue of claim 1 , wherein R 3 is
wherein R 7 is substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; and R 8 is substituted or unsubstituted alkyl.
8 . The cell or tissue of claim 1 , wherein R 3 comprises an allyl moiety.
9 . The cell or tissue of claim 1 , wherein the detectable moiety comprises a fluorescent dye.
10 . The cell or tissue of claim 1 , wherein the detectable moiety comprises streptavidin moiety.
11 . The cell or tissue of claim 1 , wherein the detectable moiety a streptavidin-biotin complex.
12 . The cell or tissue of claim 1 , wherein R 5 and R 6 are unsubstituted alkyl or unsubstituted cycloalkyl.
13 . The cell or tissue of claim 1 , wherein R 5 and R 6 are independently substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.
14 . A method of sequencing a nucleic acid in a cell or tissue comprising:
(i) incorporating in series with a nucleic acid polymerase one of four different compounds into a primer to create an extension strand, wherein said primer is hybridized to said nucleic acid and wherein each of the four different compounds comprises a unique detectable label;
(ii) detecting said unique detectable label of each incorporated compound, so as to thereby identify each incorporated compound in said extension strand, thereby sequencing the nucleic acid;
(iii) wherein each of said four different compounds is a compound of claim 1 .
15 . A method of sequencing nucleic acid in a cell or tissue comprising:
(i) providing a nucleic acid template hybridized to a primer;
(ii) extending the primer hybridized to said nucleic acid template with a compound of claim 1 ; and
(iii) identifying the compound, so as to sequence the nucleic acid.
16 . A cell or tissue comprising a compound having the formula:
wherein
R 9 and R 10 are independently a protein, bioconjugate reactive moiety, nucleotide, therapeutic moiety, nucleic acid, or a detectable moiety;
L 1 and L 2 are independently a divalent linker;
R 5 and R 6 are independently unsubstituted alkyl, —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SO 3 H, —SO 4 H, —SO 2 NH 2 , —NHNH 2 , —ONH 2 , substituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and;
W 1 and W 2 are —O—.
17 . The cell or tissue of claim 16 , wherein R 9 is a protein and R 10 is a detectable moiety.
18 . The cell or tissue of claim 16 , wherein R 9 is a nucleotide and R 10 is a detectable moiety.
19 . The cell or tissue of claim 16 , wherein R 9 is a nucleotide is a nucleotide and R 10 is a bioconjugate reactive moiety comprising a streptavidin-biotin complex.
20 . The cell or tissue of claim 16 , wherein the detectable moiety is a fluorescent dye.