Poster Presentation ASSCR, AGCTS, ISCT ANZ and Friends Joint Scientific Conference 2019

Combinations of Promoters and Virus Capsids May Affect Gene Transduction Efficiency in Adeno-associated Virus (#219)

Yoshihide YS Sehara 1 , Kuniko Shimazaki 2 , Masashi Urabe 1 , Kensuke Kawai 2 , Hiroaki Mizukami 1
  1. Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical University, Shimotsuke, Tochigi, Japan
  2. Department of Neurosurgery, Jichi Medical University, Shimotsuke, Tochigi, Japan

[Purpose] Adeno-associated virus (AAV) vector has been widely used to transduce gene of interest for its safety and efficacy. Although the most of the combinations of promoters and viral capsids are appropriate for focal gene transduction, much is unknown about the mechanism of infection. Here, we compare the gene expression levels using some combinations of promoters (cytomegalovirus (CMV), CMV immediate-early enhancer (CAG), and Synapsin 1 (Syn1)) and viral capsids (AAV5 and AAVrh10). [Method] After six combinations of AAV vectorsuch as AAV5-CMV, AAV5-CAG, AAV5-Syn1, AAVrh10-CMV, AAVrh10-CAG, and AAVrh10-Syn1 that expressed green fluorescent protein (GFP) were made, 1×1010 vg of each vector was stereotaxically administered into the right hippocampus of 4-week-old male gerbils (n = 3, each). Three weeks after injection, animals were killed for histological analysis. The GFP expressing area of three coronal sections at 1.7, 2.0, and 2.3 mm caudal to the bregma was quantified. [Results] First, AAVrh10-CMV and AAVrh10-CAG showed the widest expression of GFP (AAVrh10-CMV: 7.1 ± 1.1 mm2, AAVrh10-CAG: 6.0 ± 0.5 mm2, p < 0.001, for both, compared with control). The second widest expression of GFP was found in the AAVrh10-Syn1 and AAV5-CMV groups (AAVrh10-Syn1: 4.2 ± 0.2 mm2, AAV5-CMV: 4.2 ± 0.2 mm2, p < 0.05, for both, compared with the control). The least expression was found in the AAV-Syn1 group (AAV5-Syn1: 0.2 ± 0.2 mm2, not significant, compared with the control).  [Conclusion] Although the combinations examined in this study is limited, we should note that some of the combinations of promoters and viral capsids may lead to the failure of gene transduction.

  • Have you presented your abstract at another international meeting?: No