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US 2018/0148764 A1Published application
Enzymatic modification of nucleic acids
AssigneeThe Regents of the University of California
InventorsNeal K. Devaraj, Seth C. Alexander
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Abstract
There are provided, inter alia, methods and reagents for labeling nucleic acids.
Claims44
A method for labeling a nucleic acid, the method comprising:
- contacting the nucleic acid comprising native nucleobases with a transglycosylase and a nucleoside derivative under conditions suitable to exchange at least one the native nucleobases for the nucleoside derivative, wherein the nucleic acid is an RNA or DNA.
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A compound with structure of Formula (I):
- wherein
- L1 is a bond, —C(O)—, —C(O)O—, —C(O)NR2—, —O—, —S(O)n—, —S(O)NR2—, —C(O)NR2S(O)2—, substituted or unsubstituted alkylene, or substituted or unsubstituted heteroalkylene, substituted or unsub stituted cycloalkylene, substituted or unsub stituted heterocycloalkylene, substituted or unsubstituted aryl ene, or substituted or unsubstituted heteroaryl ene;
- R1 is substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl,
- R2 is independently hydrogen, halogen, —N3, —NO2, —CF3, —CCl3, —CBr3, —CI3, —CN, —OH, —NH2, —COOH, —CONH2, —NO2, —SH, —SO2Cl, —SO3H, —SO2Ph, —SO2NH2, —NHNH2, —ONH2, —OCH3, —NHCNHNH2, 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, and n is 0, 1, or 2.
1 more element
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A compound of structural Formula II:
- or a salt thereof,
- wherein:
- L1 is a bond, substituted or unsubstituted alkylene or substituted or unsubstituted heteroalkylene, a detectable moiety, —CX1.13, —CHX1.12, —CH2X1.1, —CN, —SOn1R1A, —SOv1NR1BR1C, —NHNR1BR1C, —ONR1BR1C, —NHC(O)NHNR1BR1C, —NHC(O)NR1BR1C, —N(O)m1, —NR1BR1C, —C(O)R1D, —C(O)OR1D, —C(O)NR1BR1C, —OR1A, —NR1BSO2R1A, —NR1BC(O)R1D, —NR1BC(O)R1D, —NR1BOR1D, —OCX1.13, —OCHX1.12, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, or substituted or unsubstituted heteroarylene;
- L2 is a bond, substituted or unsubstituted alkylene or substituted or unsubstituted heteroalkylene, a detectable moiety, —CX2.13, —CHX2.12, —CH2X2.1, —CN, —SOn1R2A,
- —SOv1NR2BR2C, —NHNR2BR2C, —ONR2BR2C, —NHC(O)NHNR2BR2C, —NHC(O)NR2BR2C, —N(O)m1, —NR2BR2D, —C(O)R2D, —C(O)OR2D, —C(O)NR2BR2C, —OR2A, —NR2BSO2R2A, —NR2BC(O)R2D, —NR2BC(O)OR2D, —NR2BOR2D, OCX2.13, —OCHX2.12, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, or substituted or unsubstituted heteroarylene;
- n1 is an integer from 0 to 4;
- m1 and v1 are independently 1 or 2;
- R3 is a detectable moiety, a biomolecule, hydrogen, halogen, —CX3.13, —CHX3.12, —CH2X3.1, —CN, —SOn1R3A,
- —SOv1NR3BR3C, NHNR3BR3C, —ONR3BR3C, —NHC(O)NHNR3BR3C, —NHC(O)NR3BR3C, —N(O)m1, —NR3BR3C, —C(O)R3D, —C(O)OR3D, —C(O)NR3BR3C, OR3A, —NR3BSO2R3A, —NR3BC(O)R3D, —NR3BC(O)OR3D, —NR3BOR3D, —OCX3.13, —OCHX3.12, 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;
- R4 is hydrogen or a ribose, wherein the ribose is part of a modified or unmodified RNA molecule, and further wherein the ribose comprises a guanine;
- R1A, R1B, R1C, R1D, R2A, R2B, R2C, R3D, R3A, R3B, R3C and R3D are independently hydrogen, halogen, —CF3, —CCl3, —CBr3, —CI3, —OH, —NH2, —COOH, —CONH2, —NO2, —SH, —SO3H, —SO4H, —SO2NH2, —NHNH2, —NHC(O)NHNH2, —NHC(O)NH2, —NHSO2H, —NHC(O)H, —NHC(O)—OH, —NHOH, —OCF3, —OCCl3, —OCBr3, —OCI3, —OCHF2, —OCHCl2, —OCHBr2, —OCHI2, 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;
- R1B, R1C, R2B, R2C, R3B and R1C substituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl;
- and
- X1.1, X2.1 and X3.1 are independently —Cl, —Br, —I or —F.
11 more elements
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A method of substituting a guanine with a PreQ1 analog within an RNA molecule, comprising
- contacting a PreQ1 analog with an RNA molecule in the presence of a transglycosylase;
- and
- (ii) allowing the transglycosylase to substitute a guanine moiety from a guanine within the RNA sequence with the PreQ1 analog thereby forming a modified RNA molecule.
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A method of screening for a transglycosylase inhibitor comprising:
- contacting a detectable PreQ1 with an RNA molecule in the presence of a transglycosylase and a test transglycosylase inhibitor;
- and
- (ii) determining whether the transglycosylase substitutes a guanine moiety from a guanine within the RNA molecule with the detectable PreQ1 analog.
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Dependencies are inferred from the claim text.