Efficient DNA–Polymer Coupling in Organic Solvents: A Survey of Amide Coupling, Thiol-Ene and Tetrazine–Norbornene Chemistries Applied to Conjugation of Poly(N-Isopropylacrylamide)

Abstract

A range of chemistries were explored for the efficient covalent conjugation of DNA to poly(N-isopropylacrylamide) (poly(NIPAM)) in organic solvents. Amide coupling and thiol–ene Michael addition were found to be ineffective for the synthesis of the desired products. However, the inverse electron-demand Diels–Alder (DAinv) reaction between tetrazine (Tz) and norbornene (Nb) was found to give DNA–polymer conjugates in good yields (up to 40%) in organic solvents (N,N-dimethylformamide, N,N-dimethylacetamide and N-methyl-2-pyrrolidone), and without the need for a catalyst. Methods for the synthesis of Tz-and Nb- functionalised DNA were developed, along with a post-polymerisation functionalisation strategy for the production of Tz-functionalised polymers.

DNA-Functionalized Dye-Loaded Polymeric Nanoparticles: Ultrabright FRET Platform for Amplified Detection of Nucleic Acids

“…However, small PMMA-MA-based NPs bearing nucleic acids have not been reported to date. The problem is inefficiency of the direct coupling of hydrophobic polymers (including PMMA-MA derivatives) with oligonucleotides because of difficulty to solubilize together highly apolar and polar species, so that solid phase synthesis is required. , However, the examples of direct modification of polymeric NPs by oligonucleotides are limited to block copolymers containing relatively large hydrophilic units with PEG linkers, which is incompatible with our FRET strategy. On the basis of our recent work, showing that a single charge per polymer is sufficient to assemble small NPs by nanoprecipitation, we expect that addition of a reactive group next to this charged group will enable preparation of functionalizable nanoantennas.…”

Synthetic approaches for copolymers containing nucleic acids and analogues: challenges and opportunities

“…10,14,15 Therefore, highly hydrophobic, nonpolar polymers are generally thought to be incompatible for conjugation with DNA, although extensive optimizations have achieved reasonable yields. 16 With regard to the bioconjugation reaction, amidation is among the first which is studied. Park and co-workers coupled poly(D,L-lactic-co-glycolic acid) (PLGA) with an antisense oligonucleotide to form an amphiphilic diblock copolymer, which self-assembled into micelles in aqueous buffer.…”

Scalable One-Pot-Liquid-Phase Oligonucleotide Synthesis for Model Network Hydrogels

“…Whereas the attachment of small molecules to DNA can be achieved using traditional post-conjugation techniques, [55][56] insufficient reactivity between end-functionalized DNA and reactive star-PEGs (or other polymers) makes starPEG-DNA conjugates difficult to obtain in high purity, definition, and large scale. [57][58][59] However, such well-defined star-DNA polymers would be interesting components for fundamental sciences and applications. For instance, star polymers with defined end-group connectivity facilitate the construction of model network materials, and are highly sought after for the topological design of network materials.…”