2.1.1 An adeno-associated virus (AAV) comprises a linear single-stranded DNA (ssDNA) genome of approximately 4.7 kb, enclosed in an icosahedron-shaped capsid consisting of three proteins.[39] The genome consists of two 145 nucleotide Long inverted terminal repeats (ITRs) at the termini, which encompass four replication (rep) and the three capsid (cap) genes. The full, non-enveloped virus is about 20 nm in diameter. As the name implies, the wild-type virus cannot replicate without the aid of additional genes provided by co-infection of adenovirus and is considered non-pathogenic. These features lend AAV to development as a delivery vector for genes that will fit into the capsid (limited to ~5 kb), and with a lower risk of infection caused by contaminating wild-type virus.[39]
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2.2.1 The most common lentiviral vectors used today are based on human immunodeficiency virus-1 (HIV-1). Native HIV-1 consists of an asymmetric cone-shaped protein capsid that is 100–200 nm long and 45–50 nm wide.[41] The capsid is surrounded by an envelope[42] and encases the 9.8 kb positive-sense RNA genome. The native HIV envelope is derived from the host cell membrane and studded with viral Env proteins.[43]
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2.3.1 Both lentiviral- and AAV-based vectors are typically manufactured by transiently transfecting mammalian cells, such as HEK293, with either three (AAV) or four (lentivirus) plasmids. AAV is typically dosed in much higher quantities for clinical use, but the common vectors for lentiviral production are also needed in large amounts.
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2.4.1 To approximate the clinical requirements for plasmid DNA, we analyzed the clinical viral vector landscape. At the end of 2023, the Alliance for Regenerative Medicine (ARM) reported (from GlobalData) 2,120 clinical trials for advanced therapies underway around the world — a decrease of 4% from 2,220 trials in 2022. The global distribution of clinical trials around the world retained a similar pattern as in 2023, with 45% in North America, 35% in Asia–Pacific, and 20% in Europe (Figure 4). The trials are being sponsored by 2,526 cell and gene therapy developers.[1]
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