Oligonucleotides: Molecular Versions for Optimal use in Vivo

Saison-Behmoaras, E.; Aerschot, Arthur Van; Duroux, Isabelle; Hendrix, Chris; Hélène, Claude; Herdewijn, Piet

doi:10.1007/978-1-4615-6405-8_4

Cited by 1 publication

(1 citation statement)

References 6 publications

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“…While xenobiotics are normally taken up by passive diffusion, considering the size of peptide-BSA-gold conjugates, passive diffusion is an unlikely translocation mechanism here. Pinocytosis and receptor-mediated endocytosis (RME) are the prime mechanisms for the uptake of large molecular structures (28). Cellular uptake of nanoparticles coated with peptide-BSA conjugates could occur by RME, or for the Tat RKKRRQRRR sequence, via the less understood peptide transduction domain mechanism (PTD).…”

Section: Preparation Of Gold Nanoparticle Vectorsmentioning

confidence: 99%

Cellular Trajectories of Peptide-Modified Gold Particle Complexes: Comparison of Nuclear Localization Signals and Peptide Transduction Domains

et al. 2004

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Gold nanoparticles modified with nuclear localization peptides were synthesized and evaluated for their subcellular distribution in HeLa human cervical epithelium cells, 3T3/NIH murine fibroblastoma cells, and HepG2 human hepatocarcinoma cells. Video-enhanced color differential interference contrast microscopy and transmission electron microscopy indicated that transport of nanoparticles into the cytoplasm and nucleus depends on peptide sequence and cell line. Recently, the ability of certain peptides, called protein transduction domains (PTDs), to transclocate cell and nuclear membranes in a receptor- and temperature-independent manner has been questioned (see for example, Lundberg, M.; Wikstrom, S.; Johansson, M. (2003) Mol. Ther. 8, 143-150). We have evaluated the cellular trajectory of gold nanoparticles carrying the PTD from HIV Tat protein. Our observations were that (1) the conjugates did not enter the nucleus of 3T3/NIH or HepG2 cells, and (2) cellular uptake of Tat PTD peptide-gold nanoparticle conjugates was temperature dependent, suggesting an endosomal pathway of uptake. Gold nanoparticles modified with the adenovirus nuclear localization signal and the integrin binding domain also entered cells via an energy-dependent mechanism, but in contrast to the Tat PTD, these signals triggered nuclear uptake of nanoparticles in HeLa and HepG2 cell lines.

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Section: Preparation Of Gold Nanoparticle Vectorsmentioning

confidence: 99%

Cellular Trajectories of Peptide-Modified Gold Particle Complexes: Comparison of Nuclear Localization Signals and Peptide Transduction Domains

et al. 2004

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Analysis of antisense oligonucleotides by on‐capillary isotachophoresis and capillary polymer sieving electrophoresis

et al. 1998

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An attempt was made to evaluate the stability of an antisense oligonucleotide against nucleases present in HBL 100ras cells. To detect nanomolar concentrations of the oligonucleotide, a sensitive detection system was required. A combination of capillary electrophoresis/laser-induced fluorescence (CE-LIF) with fluorescence derivatization did not improve the sensitivity significantly and also resulted in loss of separation of the derivatized sample. On-column isotachophoresis for the preconcentration of oligonucleotide samples in DB-17 coated capillaries filled with hydroxyethyl cellulose solution could be an alternative. The isotachophoresis (ITP) step allows injection of up to 40% of the capillary volume without loss in peak resolution and peak efficiency. Using ITP-capillary polymer sieving electrophoresis (CPSE), the limit of quantitation at a signal-to-noise ratio of 10 was 73 ng/mL for a 12-mer oligonucleotide. Using these conditions, the gain in sensitivity was 125.

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Oligonucleotides: Molecular Versions for Optimal use in Vivo

Cited by 1 publication

References 6 publications

Cellular Trajectories of Peptide-Modified Gold Particle Complexes: Comparison of Nuclear Localization Signals and Peptide Transduction Domains

Cellular Trajectories of Peptide-Modified Gold Particle Complexes: Comparison of Nuclear Localization Signals and Peptide Transduction Domains

Analysis of antisense oligonucleotides by on‐capillary isotachophoresis and capillary polymer sieving electrophoresis

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