Synthetic Virus-Derived Nanosystems (SVNs) for Delivery and Precision Docking of Large Multifunctional DNA Circuitry in Mammalian Cells.

Francesco Aulicino,Imre Berger,Julien Capin

doi:10.3390/pharmaceutics12080759

Francesco Aulicino, Imre Berger + Show 1 more

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https://doi.org/10.3390/pharmaceutics12080759

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DNA delivery is at the forefront of current research efforts in gene therapy and synthetic biology. Viral vectors have traditionally dominated the field; however, nonviral delivery systems are increasingly gaining traction. Baculoviruses are arthropod-specific viruses that can be easily engineered and repurposed to accommodate and deliver large sequences of exogenous DNA into mammalian cells, tissues, or ultimately organisms. These synthetic virus-derived nanosystems (SVNs) are safe, readily customized, and can be manufactured at scale. By implementing clustered regularly interspaced palindromic repeats (CRISPR) associated protein (CRISPR/Cas) modalities into this system, we developed SVNs capable of inserting complex DNAs into genomes, at base pair precision. We anticipate a major role for SVNs as an attractive alternative to viral vectors in accelerating genome engineering and gene therapy applications in the future.

Highlights

Programmable DNA nucleases such as clustered regularly interspaced palindromic repeats (CRISPR) associated protein 9 (CRISPR/Cas9) have enormously contributed to rejuvenate and democratize the gene editing and gene therapy fields thanks to their efficiency, cost-effectiveness, and specificity, in a variety of model organisms in ex vivo and in vivo
CRISPR system was originally discovered in bacteria as a previously unknown adaptive immune response pathway, it was successfully repurposed into a gene editing tool when Cas9, an RNA guided DNA programmable nuclease, was shown to efficiently work in mammalian cells to produce double stranded DNA breaks (DSBs) at desired genomic loci with very high efficiency and little off-target effects [1,2]
According to the subsequent DNA repair mechanism, Cas9-induced DSBs can be resolved with the production of small insertion/deletions through the non-homologous end joining (NHEJ) DNA repair pathway, or precise gene editing outcomes when the homology-directed repair (HDR) pathway is exploited in combination with a suitable DNA template [2]

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Programmable DNA nucleases such as clustered regularly interspaced palindromic repeats (CRISPR) associated protein 9 (CRISPR/Cas9) have enormously contributed to rejuvenate and democratize the gene editing and gene therapy fields thanks to their efficiency, cost-effectiveness, and specificity, in a variety of model organisms in ex vivo and in vivo. CRISPR system was originally discovered in bacteria as a previously unknown adaptive immune response pathway, it was successfully repurposed into a gene editing tool when Cas9, an RNA guided DNA programmable nuclease, was shown to efficiently work in mammalian cells to produce double stranded DNA breaks (DSBs) at desired genomic loci with very high efficiency and little off-target effects [1,2].

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