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Adenoviral Vector Production and Troubleshooting

Posted by Karen Guerin on Jun 14, 2018 7:44:26 AM

Adenoviral vectors (AdV) are attractive vectors for research applications and gene therapy: they can be produced at high titers, can accommodate large transgenes, transduce quiescent and dividing cells, and do not integrate into the host’s genome. The main challenge with using AdV is that it triggers a strong immune response after in vivo administration, which results in the death of transduced cells and loss of transgene expression (Interestingly, the strong immunogenicity of AdVs is what makes them ideal candidates for applications in oncolysis and vaccination!)

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

Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

Posted by Kendall Morgan on Sep 30, 2014 4:50:49 PM

Researchers have shown that it is possible to deliver RNA-guided CRISPR/Cas9 nuclease complexes  using adenoviral vectors (AdVs), to a wide range of human cells, including mesenchymal stem cells, and in a rather straightforward manner. These adenoviral CRISPR/Cas9 genome editing tools developed and demonstrated by Manuel Gonçalves and his colleagues at Leiden University Medical Center are now available at Addgene along with a description of their experimental protocol. The three plasmids which have been deposited to Addgene are: pAdSh.PGK.Cas9, pAdSh.U6.gRNAS1, pAdSh.U6.gRNAGFP.

“Although AdVs are being deployed for delivering zinc-finger nucleases into human cells, we think they are still underused in the emerging field of genome editing,” Gonçalves said. “In contrast, AdVs are extensively being explored for genetic vaccination and oncolytic approaches. In genome editing, they are not used much, but we do think they have a very bright future.”

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Topics: CRISPR, CRISPR Expression Systems and Delivery Methods, Adenoviral Vectors

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