Publications by authors named "Daniela Goldnau"
Adeno-associated virus type 2 (AAV-2) targeting vectors have been generated by insertion of ligand peptides into the viral capsid at amino acid position 587. This procedure ablates binding of heparan sulfate proteoglycan (HSPG), AAV-2's primary receptor, in some but not all mutants. Using an AAV-2 display library, we investigated molecular mechanisms responsible for this phenotype, demonstrating that peptides containing a net negative charge are prone to confer an HSPG nonbinding phenotype.
View Article and Find Full Text PDFBackground: Viruses are being exploited as vectors to deliver therapeutic genetic information into target cells. The success of this approach will depend on the ability to overcome current limitations, especially in terms of safety and efficiency, through molecular engineering of the viral particles.
Methods: Here we show that in vitro directed evolution can be successfully performed to randomize the viral capsid by error prone PCR and to obtain mutants with improved phenotype.
Article Synopsis
- Researchers genetically modified adeno-associated virus (AAV) to include enhanced green fluorescent protein (GFP) to visualize viral trafficking and observe how AAV enters the nucleus.
- In cells infected only with AAV, viral capsids were slow to enter the nucleus, remaining mostly in the perinuclear area for several hours, while co-infection with adenovirus 5 (Ad5) significantly accelerated their nuclear entry.
- The study concludes that intact AAV capsids do not efficiently enter the nucleus, suggesting viral uncoating occurs prior to or during the entry process instead.