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Viral Vectors 101: Parts of the AAV Packaging Plasmid

Posted by Beth Kenkel on Dec 3, 2020 9:15:00 AM

Your next cool experiment requires some AAV.  Where do you start? Plasmids of course! You just need three plasmids to start making the AAVs you need for your experiment: 

  1. the packaging plasmid which contains the AAV structural and packaging genes,
  2. the adenoviral helper plasmid which contains the proteins needed for the virus to replicate, 
  3. and the transfer plasmid which contains the viral genome.

In today’s blog post, we’ll focus on the packaging plasmid and take a look at each of its parts.

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Topics: Viral Vectors, Viral Vectors 101, AAV

Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

Posted by Beth Kenkel on Sep 1, 2020 9:15:00 AM

Adeno-associated virus (AAV) has many features which make it a great viral vector, but its packaging capacity is limited to ~4.7kb, or roughly half the packaging limits of lentiviral and adenoviral vectors. While many transgene will fit within this limit, some like prime editing's PE2 enzyme do not. So how do you fit a big gene into a tiny vector like AAV? By breaking the transgene into smaller pieces.

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Topics: Viral Vectors, AAV

Viral Vectors 101: Parts of the AAV Transfer Plasmid

Posted by Beth Kenkel on Aug 11, 2020 9:15:00 AM

So you have this awesome experiment you want to do, but it requires some AAV. You’ve never worked with AAV before, but you aren’t going to let that stop you. Where do you start? Turns out like all good experiments, making AAV starts with some plasmids. You just need three plasmids to start making AAV

  1. the packaging plasmid which contains the structural and packaging genes,
  2. the adenoviral helper plasmid which contains the proteins needed for the virus to replicate, 
  3. and the transfer plasmid which contains the viral genome.

In today’s blog post, we’ll focus on the AAV transfer plasmid and take a look at each of its parts.

Read More >

Topics: Viral Vectors, Viral Vectors 101, AAV

Viral Genome Sequencing - A One Stop Shop for AAV Identity and Serotype Confirmation

Posted by Meghan Rego on Apr 23, 2020 9:15:00 AM

One of the main tenets of our quality control (QC) mantra is to do our utmost to ensure that scientists are receiving the exact materials that they think they are. To address this, several years ago we partnered with Seqwell to establish a next-generation sequencing (NGS) QC platform that allows us to provide scientists with complete plasmid sequences. After launching our viral service in 2016, we adapted this platform to accommodate adeno-associated viral vector (AAV) samples and created a simplified process, termed viral genome sequencing (VGS), to confirm the identity and serotype of our AAV preparations. We recently published a description of the VGS process and use cases in Human Gene Therapy.

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Topics: Viral Vectors, Viral Vector Protocols and Tips, AAV

Synthetic promoter AAVs for cell-type specific expression in retinal cells

Posted by Beth Kenkel on Mar 12, 2020 9:15:00 AM

Adeno-associated virus (AAV) is a popular tool for gene delivery, but it has a drawback: how do you ensure your gene goes where you want it to? Knowing that a gene is expressed in a particular cell type is important not only for translational research, such as gene therapy, but also basic research. To improve cell-type specificity of AAV, work has focused on modifying the outside protein shell, or capsid, of the virus so that it only enters and delivers it cargo to certain cell types. 

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Topics: Viral Vectors, Cell Tracing, AAV

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