Gene Therapy Vectors Based on Adeno-Associated Virus Type 1

Abstract

The complete sequence of adeno-associated virus type 1 (AAV-1) was defined. Its genome of 4,718 nucleotides demonstrates high homology with those of other AAV serotypes, including AAV-6, which appears to have arisen from homologous recombination between AAV-1 and AAV-2. Analysis of sera from nonhuman and human primates for neutralizing antibodies (NAB) against AAV-1 and AAV-2 revealed the following. (i) NAB to AAV-1 are more common than NAB to AAV-2 in nonhuman primates, while the reverse is true in humans; and (ii) sera from 36% of nonhuman primates neutralized AAV-1 but not AAV-2, while sera from 8% of humans neutralized AAV-2 but not AAV-1. An infectious clone of AAV-1 was isolated from a replicated monomer form, and vectors were created with AAV-2 inverted terminal repeats and AAV-1 Rep and Cap functions. Both AAV-1- and AAV-2-based vectors transduced murine liver and muscle in vivo; AAV-1 was more efficient for muscle, while AAV-2 transduced liver more efficiently. Strong NAB responses were detected for each vector administered to murine skeletal muscle; these responses prevented readministration of the same serotype but did not substantially cross-neutralize the other serotype. Similar results were observed in the context of liver-directed gene transfer, except for a significant, but incomplete, neutralization of AAV-1 from a previous treatment with AAV-2. Vectors based on AAV-1 may be preferred in some applications of human gene therapy.

Nutrient‐sensitive transcription factors TFEB and TFE 3 couple autophagy and metabolism to the peripheral clock

“…pAAV2/1-CMV-EGFP, pAAV2/1-CMV-TFE3 (Pastore et al, 2017), and pAAV2/1-CMV-mTFEB-3xFlag (Mansueto et al, 2017) were triple-transfected in subconfluent 293 cells along with pAd-Helper and pack2/1 packaging plasmids as previously described (Xiao et al, 1999). Recombinant vectors were purified by two rounds of CsCl gradient centrifugation, as previously described (Xiao et al, 1999). Vector titers, expressed as genome copies per milliliter (GC/ml), were assessed by both PCR quantification and dot-blot analysis.…”

Humoral Immunity to AAV-6, 8, and 9 in Normal and Dystrophic Dogs

“…15.1 1.3 -1.2 1.0 AAV-6 was originally isolated from an adenovirus stock (Rutledge et al, 1998). However, it may actually represent a natural recombinant of AAV-1 and 2, because its first 508 nucleotides are identical to AAV-2 and the remaining nucleotides are identical to AAV-1 (Xiao et al, 1999). AAV-1 and 2 were also isolated from adenovirus stocks (Atchison et al, 1965;Xiao et al, 1999).…”

“…However, it may actually represent a natural recombinant of AAV-1 and 2, because its first 508 nucleotides are identical to AAV-2 and the remaining nucleotides are identical to AAV-1 (Xiao et al, 1999). AAV-1 and 2 were also isolated from adenovirus stocks (Atchison et al, 1965;Xiao et al, 1999). AAV-8 and 9 were isolated by PCR from rhesus monkey and human tissues, respectively (Gao et al, 2002(Gao et al, , 2004.…”

Production of adeno-associated virus (AAV) serotypes by transient transfection of HEK293 cell suspension cultures for gene delivery

“…However, the most commonly studied are the eleven AAV serotypes (AAV1 to AAV11) isolated from human, simian, and rhesus and cynomolgus monkeys. Serotype AAV6 (Rutledge et al, 1998;Xiao et al, 1999) is believed to be a recombinant product of AAV2 and AAV1. AAV serotype 2 has been extensively studied for delivering genes to treat a variety of diseases.…”