Templated DNA Cross‐Linking: Towards a Non‐Invasive Singlet‐Oxygen‐Based Triggering Method

Abstract

Singlet oxygen-induced DNA interstrand crosslinking was achieved at intracellularly relevant concentrations using a templated reaction setup. The target oligonucleotide serves as a template ensuring proximity between a "furan warhead"-containing probe and an activator probe containing a photosensitizer able to generate 1 O2. Visible light irradiation of this self-activating assembly allows to achieve highly selective interstrand crosslinking at concentration levels relevant for intracellular applications.

A Photosensitized Singlet Oxygen (1O2) Toolbox for Bio-Organic Applications: Tailoring 1O2 Generation for DNA and Protein Labelling, Targeting and Biosensing

“…In a follow-up of this study, furan activation was then achieved by 1 O 2 -mediated photo-oxidation, in presence of MB as PS [ 172 ], and later on extended from DNA to RNA [ 173 ]. The technology was further successfully applied in a DNA-templated fashion, upon insertion of the PS in another DNA strand, adjacently hybridized to the target [ 174 , 175 ] ( Figure 12 B). A similar concept of exploiting DNA as template, was applied through the co-localization of MB, a weak G-Quadruplex (G4) binder, with a furan-containing G4-ligand, in order to enable selective alkylation of G4-DNA over dsDNA, triggered by red light [ 176 ].…”

“…In this case, the use of two separate strands, one bearing furan and the other one bearing the required PS, both adjacently hybridized to the target strand, effectively leads to formation of ICL products. On the other hand, the introduction of both moieties on the same probe, led to the degradation of the targeting probe, with no ICL formation [ 175 , 301 ].…”

“…( A ) Original methodology proposed by Madder in [ 172 ]. ( B ) Follow-up study with templated DNA-CL described by Madder and colleagues in refs [ 174 , 175 ]. ( C ) Protein–RNA CL upon incorporation of genetically encoded furan moiety.…”

Red Light-Initiated Cross-Linking of NIR Probes to Cytoplasmic RNA: An Innovative Strategy for Prolonged Imaging and Unexpected Tumor Suppression

“…Unfortunately, the broad biological applications of this photo-cross-linking approach in living systems were impeded by the limited tissue penetration and potential phototoxicity of ultraviolet (UV) light . Madder and co-workers have recently developed a versatile red light-controlled cross-linking chemistry based on furan for studying a large variety of biomolecular interactions with spatiotemporal resolution, including DNA–DNA , and protein–RNA. , However, in vivo application of this cross-linking strategy has not been exploited yet. Ribonucleic acid (RNA) as a central conduit plays a pervasive and important role in cell information transmission and gene regulation .…”

Visible-light triggered templated ligation on surface using furan-modified PNAs

“…53 Furan oxidation was initially triggered by using N-bromosuccinimide, but was later accomplished in a visible light-triggered manner by exploiting in situ singlet oxygen production via photosensitizer (PS) induced activation. [54][55][56] This methodology was subsequently applied as a means to immobilize DNA on surfaces, 57 and later on extended to the context of peptide labelling and peptide-peptide ligation, exploiting in the latter cases the chemo-selective reactivity of the activated furan moiety towards a-effect nucleophiles. 58,59 In this work the rst example of a light-triggered templated ligation exploiting a stable but pro-reactive furan moiety which can be oxidized to its reactive keto-enal form by light induced singlet oxygen ( 1 O 2 ) generation (Fig.…”