Compositions and methods for modifying dna
AssigneeThe Regents of the University of California
InventorsEmber M. Tota, Neal Devaraj
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Abstract
Provided herein are, inter alia, compositions and methods for modifying a DNA molecule. The methods include contacting a DNA molecule with a transglycosylase enzyme and a Pre-queuosinel (preQi) analog or derivative, wherein a guanine nucleobase within a hairpin structure in the DNA molecule is exchanged for the preQi analog or derivative.
Claims54
A method of modifying a DNA molecule, the method comprising contacting the DNA molecule with a tRNA-guanine transglycosylase (TGT) enzyme and a PreQ1 analog,
- wherein the DNA molecule comprises a guanine nucleobase within a loop portion of a hairpin in the DNA molecule, and
- wherein the PreQ1 analog has the formula:
- wherein Q is not hydrogen.
A method of substituting a guanine nucleobase within a DNA molecule with a PreQ1 analog, the method comprising contacting the DNA molecule and PreQ1 analog with a tRNA-guanine transglycosylase (TGT) enzyme,
- wherein the guanine nucleobase is within a loop portion of a hairpin in the DNA molecule, and
- wherein the PreQ1 analog has the formula:
- wherein Q is not hydrogen.
A DNA compound having the formula:
- wherein
- R1 is hydrogen, a deoxynucleotide, or a first DNA sequence comprising the 5′ of a DNA hairpin;
- R2 is hydrogen, a deoxynucleotide, or a second DNA sequence comprising the 3′ of the DNA hairpin;
- L1 and L2 are independently a bond, —O—, —S—, —NH—, —C(O)NH2, —NHC(O)—, —C(O)O—, —OC(O)—, —S(O)2NH2, —NHS(O)2—, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, or substituted or unsubstituted heteroarylene, a peptide linker, or a cleavable linker;
- R3 is a detectable moiety, a biomolecular moiety, a therapeutic moiety, 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;
- R3A, R3B, R3C and R3D are independently hydrogen, halogen, —CF3, —CCl3, —CBr3, —CI3, —CHCl2, —CHBr2, —CHF2, —CHI2, —CH2Cl, —CH2Br, —CH2F, —CH2I, —CN, —OH, —NH2, —COOH, —CONH2, —NO2, —SH, —SO3H, —SO4H, —SO2NH2, —NHNH2, —ONH2,
- —NHC(O)NHNH2, —NHC(O)NH2, —NHSO2H, —NHC(O)H, —NHC(O)—OH, —NHOH, —OCF3, —OCCl3, —OCBr3, —OCI3, —OCHF2, —OCHCl2, —OCHBr2,
- —OCHI2, —OCH2Cl, —OCH2Br, —OCH2I, —OCH2F, 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;
- R3B and R3C substituents bonded to the same nitrogen atom may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl;
- and
- X3.1 is independently —Cl, —Br, —I or —F.
Dependencies are inferred from the claim text.