Overview

Recombinant adeno-associated virus (rAAV) is a viral vector widely used in gene therapy and molecular biology research. In addition to be non-pathogenic, rAAVs are very attractive because they can efficiently deliver genetic material (transgene) into target cells and tissues upon transduction, with a low immunogenicity.

At Polyplus, we have developed an Easy rAAV pTransgene plasmid to be co-transfected into packaging cells along with the pHelper plasmid (e.g. pPLUS® AAV-Helper), the pRepCap plasmid, and transfection reagents (e.g. FectoVIR®-AAV), enabling the efficient production of viral particles to further transduce targeted cells/tissues to express the gene of interest. Our Easy rAAV pTransgene plasmid has been designed as a single-stranded AAV (ssAAV) and contains ITR sequences from AAV2.

Genetic Features:

Bgh-PA : The Bovine growth hormone (Bgh) polyA (PA)signal allows for polyadenylation and thereby, provides transcription termination.

eZ-Ori : The e-Zyvec® Origin of replication (eZ-Ori) is a high copy number origin of replication (pMB1 Ori) optimized by e-Zyvec for optimal assembly and plasmid amplification yield in E. coli.

GOI : The gene of interest (GOI) is the coding sequence delivered into the target cells. In general, the capacity of an AAV vector is limited to approximately 4.7kb (promoter and termination site included) of DNA, as this is the maximum size that the virus can efficiently carry. In practice, it is advisable to stay below this limit to ensure optimal packaging and transduction efficiency.

ITR : Inverted Terminal Repeats (ITR) are sequences essential for the replication and packaging of the (recombinant) viral genome. They serve as recognition sites for viral enzymes, initiating the process of DNA replication and packaging into new virus particles.

KanR : The Kanamycin Resistance cassette is the prokaryotic expression cassette that will help us identify and select the conform vector candidates during the amplification process by E.coli.

Kozak : Is a sequence of nucleotides in eukaryotic mRNA that is recognized and used by the ribosome as the start site for translation.
Promoter: The promoter is the DNA sequence driving the expression of a GOI. The expression level of the GOI is cell-type and promoter-dependent. In our “easy plasmid offer”, we offer a variety of eight promoters to fit your needs.

If you require a more specific set of features for your plasmid, we provide a fully customizable plasmid construction service, do not hesitate to contact us regarding your projects.

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