[Nuclear localization of oligonucleotides decoy effect on nuclear factor-kappaB activity]

Area of Science:

• Molecular Biology
• Cell Biology
• Bioconjugation Chemistry

Context:

• Nuclear factor-kappaB (NF-kappaB) is a key transcription factor involved in inflammatory and immune responses.
• Dysregulation of NF-kappaB activation is implicated in various diseases, including cancer and autoimmune disorders.
• Targeting NF-kappaB activation is a significant therapeutic strategy.

Purpose:

• To investigate the impact of nuclear localization signal (NLS) peptide conjugation on the efficacy of decoy oligonucleotides (ODNs decoy) in inhibiting NF-kappaB activation.
• To develop a method for enhancing the nuclear delivery of ODNs decoy.
• To assess the safety and effectiveness of NLS-conjugated ODNs decoy in TNF-alpha induced HeLa cells.

Summary:

• A mercapto group-modified NLS peptide was covalently conjugated to amino group-modified NF-kappaB ODNs decoy using a Sulfo-SMCC cross-linker.
• The resulting NLS-ODNs decoy was efficiently transfected into HeLa cells, with a high nuclear localization rate of 17.9%.
• MTT assays confirmed no significant impact on cell viability, while electrophoretic mobility shift assays (EMSA) demonstrated significant inhibition of NF-kappaB activation in nuclear extracts.

Impact:

• This study presents a novel covalent conjugation strategy for NLS peptides and ODNs decoy.
• The NLS-ODNs decoy effectively enhances nuclear delivery, leading to improved inhibition of NF-kappaB activation.
• This approach offers a promising strategy for developing targeted therapies against NF-kappaB-mediated diseases.