Shortly after entering the cell, HIV-1 copies its genomic RNA into double-stranded DNA in a process known as reverse transcription. This process starts inside a core consisting of an enclosed lattice of capsid proteins that protect the viral RNA from cytosolic sensors and degradation pathways. To accomplish reverse transcription and integrate cDNA into the host cell genome, the capsid shell needs to be disassembled, or uncoated. Premature or delayed uncoating attenuates reverse transcription and blocks HIV-1 infectivity. Small molecules that bind to the capsid lattice of the HIV-1 core and either destabilize or stabilize its structure could thus function as effective HIV-1 inhibitors. To screen for such compounds, we modified our recently developed FAITH assay to allow direct assessment of the stability of in vitro preassembled HIV-1 capsid-nucleocapsid (CANC) tubular particles. This new assay is a high-throughput fluorescence method based on measuring the amount of nucleic acid released from CANC complexes under disassembly conditions. The amount of disassembled CANC particles and released nucleic acid is proportional to the fluorescence signal, from which the relative percentage of CANC stability can be calculated. We consider our assay a potentially powerful tool for in vitro screening for compounds that alter HIV disassembly.

• MeSH
• HIV infekce farmakoterapie   MeSH
• HIV-1 účinky léků   fyziologie   MeSH
• látky proti HIV chemie   izolace a purifikace   farmakologie   MeSH
• lidé MeSH
• nukleokapsida analýza   účinky léků   MeSH
• proteiny virového jádra chemie   genetika   metabolismus   MeSH
• RNA virová genetika   MeSH
• rychlé screeningové testy MeSH
• sekvence aminokyselin MeSH
• sekvence nukleotidů MeSH
• svlékání virového obalu účinky léků   genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• látky proti HIV MeSH
• proteiny virového jádra MeSH
• RNA virová MeSH

Despite successful vaccination programs and effective treatments for some viral infections, humans are still losing the battle with viruses. Persisting human pandemics, emerging and re-emerging viruses, and evolution of drug-resistant strains impose continuous search for new antiviral drugs. A combination of detailed information about the molecular organization of viruses and progress in molecular biology and computer technologies has enabled rational antivirals design. Initial step in establishing efficacy of new antivirals is based on simple methods assessing inhibition of the intended target. We provide here an overview of biochemical and cell-based assays evaluating the activity of inhibitors of clinically important viruses.

• Klíčová slova
• Assay, Assembly, Cell-based, Entry, High-throughput screening, In vitro, Inhibitor, Method, Replication, Virus,
• MeSH
• antivirové látky farmakologie   MeSH
• fyziologie virů účinky léků   MeSH
• inhibitory enzymů farmakologie   MeSH
• interakce hostitele a patogenu účinky léků   MeSH
• internalizace viru účinky léků   MeSH
• kapsida účinky léků   metabolismus   MeSH
• lidé MeSH
• preklinické hodnocení léčiv metody   MeSH
• replikace viru účinky léků   MeSH
• rychlé screeningové testy metody   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• antivirové látky MeSH
• inhibitory enzymů MeSH