How is AAV manufactured?
AAV viral vectors are produced from packaging cell lines following transfection of the AAV construct and the co-infection with a helper virus, such as adenovirus (Ad) or Herpes Simplex Virus (HSV) or via a single infection with a recombinant helper viral vector containing the rAAV genome.
What is plasmid AAV?
AAV Components These ITRs base pair to allow for synthesis of the complementary DNA strand. These genes (E4, E2a and VA) mediate AAV replication. The transfer plasmid, Rep/Cap, and the helper plasmid are transfected into HEK293 cells, which contain the adenovirus gene E1+, to produce infectious AAV particles.
How are AAVs produced?
AAV production requires cytopathogenic effects, which only occur after co-infection with a helper adenovirus or herpesvirus8,9. Furthermore, specific promoters, the Cre/flox system, and gene editing via CRISPR render AAVs cell-specific and allow novel approaches in gene therapy and animal research.
What is mini circle DNA?
Minicircle DNA (mcDNA) is a smaller and safer version of non-viral DNA vectors that results from a cutting-edge in vivo recombination process to excise prokaryotic sequences from plasmid DNA (pDNA).
Are AAV biologics?
AAV Technology – Catalent Biologics. AAV, or adeno-associated virus, is considered the gold standard for in vivo gene delivery transfer and has become the leading viral vector choice for gene therapies today.
Is AAV secreted?
AAV was found to be secreted by an unknown mechanism into cell culture media using adherent HEK293 cells and harvesting rAAV from the media at a late time-point post-transfection.
What is AAV packaging?
Packaging an infective recombinant adeno-associated virus (AAV) requires the presence of certain adenoviral gene products. The AAV Helper Free System eliminates the need for a helper adenovirus, making the system safer and more convenient.
Is AAV single or double stranded?
Adeno-associated virus (AAV) vectors package single-stranded genomes and require host-cell synthesis of the complementary strand for transduction. However, when the genome is half wild-type size, AAV can package either two copies, or dimeric inverted repeat DNA molecules.
Does AAV integrate into genome?
When AAV infects a cell alone, its gene expression is repressed (AAV does not replicate), and its genome is incorporated into the host genome (into human chromosome 19). In the absence of helper virus or genotoxic factors, AAV DNA can either integrate into the host genome or persist in episomal form.
Does AAV integrate?
AAV vectors can integrate by homologous recombination, which is termed AAV-mediated gene targeting [27]. In order to promote homologous recombination, a vector that contains homology arms derived from genomic chromosomal DNA that flank the genetic modification being introduced is used.
Which is plasmid for the production of AAV?
The pDG plasmids for the production of adeno-associated virus (AAV) form a 2-plasmid system.
Where did plasmidfactory get its license from?
DKFZ-License. PlasmidFactory has obtained an exclusive, worldwide license for the production, distribution and use of the helper & packaging plasmids of the pDG and pDP families from the German Cancer Research Centre (DKFZ).
Who is the leading contract manufacturer for plasmid DNA?
PlasmidFactory is Europe’s leading contract manufacturer and service provider for plasmid DNA. Home Products & Service Custom Manufacturing Plasmid DNA Minicircle DNA In Stock Service AAV Plasmids
How are plasmids used to produce infectious particles?
Many of these plasmids encode the AAV genome (i.e., the genetic information that will be contained in the virion) and/or can be used to generate infectious AAV particles. Additional helper plasmids (which may not be available at Addgene) may be required to produce infectious AAV particles.