A5085243352- RNA Interference and Gene Delivery
- Advanced biosensing and bioanalysis techniques
- DNA and Nucleic Acid Chemistry
- Virus-based gene therapy research
- HIV Research and Treatment
- Viral gastroenteritis research and epidemiology
- HIV/AIDS drug development and treatment
- CRISPR and Genetic Engineering
- Bacteriophages and microbial interactions
- Retinal Development and Disorders
- Lipid Membrane Structure and Behavior
- Microfluidic and Bio-sensing Technologies
- Congenital heart defects research
- Viral Infectious Diseases and Gene Expression in Insects
- Immunotherapy and Immune Responses
- Cytomegalovirus and herpesvirus research
- Molecular Biology Techniques and Applications
- Nanoparticle-Based Drug Delivery
- Animal Genetics and Reproduction
- Genetic and Kidney Cyst Diseases
- 3D Printing in Biomedical Research
- Biochemical and Structural Characterization
- Neurogenesis and neuroplasticity mechanisms
- Viral Infections and Immunology Research
- Vascular Malformations and Hemangiomas
OZ Biosciences (France)
2009-2023
Cell and Gene Therapy Catapult
2008
University of California, Irvine
2007
Genex Systems (United States)
2003
University of California, San Diego
2001
University of California, San Francisco
1996-1998
Inserm
1993-1994
Centre d’Immunologie de Marseille-Luminy
1991-1994
Centre National de la Recherche Scientifique
1991-1994
We propose a mechanism for oligonucleotide (ODN) release from cationic lipid complexes in cells that accounts various observations on lipid-nucleic acid-cell interactions. Fluorescent confocal microscopy of treated with rhodamine-labeled liposome/ fluorescein-labeled ODN (F-ODN) show the F-ODN separates after internalization and enters nucleus leaving fluorescent cytoplasmic structures. displacement complex was studied by resonance energy transfer. Anionic liposome compositions (e.g.,...
We have designed a cationic amphipathic peptide, KALA (WEAKLAKALAKALAKHLAKALAKALKACEA), that binds to DNA, destabilizes membranes, and mediates DNA transfection. undergoes pH-dependent random coil α-helical conformational change as the pH is increased from 5.0 7.5. One face displays hydrophobic leucine residues, opposite hydrophilic lysine residues. KALA-mediated release of entrapped aqueous contents neutral negatively charged liposomes increases with increasing helical content....
There are many very effective methods to introduce transcriptionally active DNA into viable cells but approaches deliver functional proteins limited. We have developed a lipid-mediated delivery system that can or other bioactive molecules living cells. This is composed of new trifluoroacetylated lipopolyamine (TFA-DODAPL) and dioleoyl phosphatidylethanolamine (DOPE). cationic formulation successfully delivered antibodies, dextran sulfates, phycobiliproteins, albumin, enzymes...
We describe an effective approach using a peptide nucleic acid (PNA) "clamp" to directly and irreversibly modify plasmid DNA, without affecting either its supercoiled conformation or ability be efficiently transcribed. To demonstrate this highly fluorescent preparation of DNA was generated by hybridizing fluorescently labeled PNA the plasmid. Fluorescent prepared in way neither functionally nor conformationally altered. binding sequence specific, saturable, extremely stable, did not...
Five analytical assays are described that provide a platform for systematically evaluating the effect of formulation variables on physical properties cationic lipid-DNA complexes (lipoplexes). The (i) lipid recovery, (ii) total DNA, (iii) free (iv) nuclease sensitivity, and (v) stability by filtration. Lipid recovery was determined measuring primary amino groups labeled with fluorescamine reagent in presence detergent Zwittergent. Zwittergent effective at disrupting lipoplexes, making amine...
Phosphodiester and phosphorothioate oligonucleo- tides in a , configurations directed against the initiation codon region of HIV-1 rev gene were evaluated for their ability to inhibit replication acutely chronically infected human CEM cells.Encapsulation antibody-targeted liposomes (immuno- liposomes) permitted intracellular delivery distinc- tion between oligonucleotide-mediated inhibition viral entry effects on RNA.Our results are consistent with four mechanisms antiviral activity these...
Advances in RNA technology during the past two decades have led to construction of replication-competent RNA, termed replicons, RepRNA, or self-amplifying mRNA, with high potential for vaccine applications. Cytosolic delivery is essential their translation and self-replication, without infectious progeny generation, providing levels antigen expression inducing humoral cellular immunity. Synthetic nanoparticle-based vehicles can both protect molecules facilitate targeting dendritic...
The use of synthetic RNA for research purposes as well RNA-based therapy and vaccination has gained increasing importance. Given the anatomical seclusion eye, small interfering (siRNA)-induced gene silencing bears great potential targeted reduction pathological expression that may allow rational treatment chronic eye diseases in future. However, there is yet an unmet need techniques providing safe efficient siRNA delivery to retina. We used magnetic nanoparticles (MNPs) force (Reverse...
Two approaches are described for stably conjugating peptides, proteins and oligonucleotides onto plasmid DNA. Both methods use a peptide nucleic acid (PNA) clamp, which binds irreversibly specifically to binding site cloned into the plasmid. The first approach uses biotin-conjugated PNA clamp that can be used introduce functional biotin groups streptavidin bind. Atomic force microscopy images of linearized show localized at predicted on DNA strand. Peptides containing free thiol were...
Primary neural stem cells (NSCs) can be cultivated and differentiated in vitro but are difficult to transfect using conventional methods. We describe a simple rapid magnetofection-based method suitable for the lab bench as well high-throughput projects. Our yields high transfection efficiency used deciphering genetic control of cell differentiation.
Antisense oligonucleotides inhibit HIV replication in vitro, but their activity is limited by sensitivity to nucleases and low cellular uptake. To see whether these problems could be circumvented, we compared effects of HIV-1 rev tat gene-specific antisense phosphodiester or phosphorothioate oligonucleotides, either free solution encapsulated antibody-targeted liposomes (immunoliposomes), on acutely chronically infected cells. Phosphodiester were inactive form cells (up a concentration 50...
Cationic lipid-nucleic acid complexes are widely used to deliver oligonucleotides, RNA and DNA into cells. Although much has been learned about the structure forces that hold complex together, an understanding of mechanism release nucleic acids from cells lacking. Recent studies have shown anionic liposomes with compositions similar cytoplasmic face endosomal membrane potent agents for inducing rapid oligonucleotides cationic complexes. Based upon these results, we propose after...
An approach is described for makingtranscriptionally active PCR (TAP) fragments that were used directly in vitro and vivo expression experiments. TAP encoding reporter genes amplified 1 day using typical methodology expressed cultured cells mice at levels comparable with a widely cytomegalovirus promoter-based plasmid vector. Following intramuscular injection, fragment hepatitis B surface antigen (HBsAg) induced anti-HBsAg antibody titers those by supercoiled the same antigen. Epitope-tagged...