Design, synthesis and use of synthetic nucleotides comprising charge mass tags

Inventors

O'Halloran, Jonathan J. • Hedley, Joseph H.

Assignees

QuantumDx Group Ltd

Abstract

Embodiments of the present disclosure relate generally to reporter compositions which are synthetic nucleotides that comprise nucleotides with a high charge mass moiety attached thereto via a linker molecule. The linker molecules can vary in length in part to enable the high charge mass moiety to extend out from a DNA polymerase complex so that polymerization may not be influenced.

Core Innovation

The invention provides a reporter composition that includes a nucleotide or its derivative, a high charge mass moiety, and a linker molecule attached between the nucleotide or its derivative and the high charge mass moiety. The high charge mass moiety comprises an aromatic or aliphatic skeleton having one or more charged groups selected from tertiary amino, carboxyl, hydroxyl, phosphate, phenolic hydroxy, and combinations thereof, where the charge mass is sufficient to generate a detectable change in a property of a sensitive detection nanostructure or nano-/micro-sensor operably coupled to the reporter composition.

The linker molecule comprises a linear or branched chain comprising one or more selected from alkyl group, oxy alkyl group, alcohol group, carboxyl group, amine group, amide group, aromatic group, and naphthalene group, and combinations thereof. The linker is chemically cleavable or photocleavable, including specific linker compounds, and may comprise variable-length and/or variable functional group count, with R comprising an alkyl group, an oxy alkyl group, hydrocarbon, a hydrazone, a peptide linker, or a combination thereof.

The disclosed charged reporter moiety is configured to protrude from a DNA/RNA polymerase complex to avoid interfering with polymerization. The reporter compositions are described for nucleotide sequencing use in a kit for determining nucleotide sequence, and the reporter charge can be positive, negative, or pH-dependent, including a net charge mass that is positive in acidic pH and negative in alkaline pH, or pH-variable behavior.

Claims Coverage

The independent claims cover reporter compositions comprising a nucleotide or its derivative, a high charge mass moiety, and a linker molecule attached between them. The consolidated claim coverage distinguishes four inventive feature groups: the charged high charge mass moiety, the linker structure and functional groups, chemically cleavable linker embodiments, and photocleavable linker embodiments.

The independent claims are unified by a nucleotide or derivative linked to a high charge mass moiety having a charge mass sufficient to produce a detectable change in a coupled sensitive detection nanostructure or nano-/micro-sensor. The claims distinguish inventive scope by specifying charged-group composition and aromatic or aliphatic skeleton for the high charge mass moiety, linker structure as linear or branched chain with defined functional groups, and whether the linker is chemically cleavable or photocleavable, including specific named cleavable linker compounds.

Stated Advantages

Documented Applications