Establishment of an AAV reverse infection-based array.
BACKGROUND
The development of a convenient high-throughput gene transduction approach is critical for biological screening. Adeno-associated virus (AAV) vectors are broadly used in gene therapy studies, yet their applications in in vitro high-throughput gene transduction are limited.


RESULTS
We established an AAV reverse infection (RI)-based method in which cells were transduced by quantified recombinant AAVs (rAAVs) pre-coated onto 96-well plates. The number of pre-coated rAAV particles and number of cells loaded per well, as well as the temperature stability of the rAAVs on the plates, were evaluated. As the first application of this method, six serotypes or hybrid serotypes of rAAVs (AAV1, AAV2, AAV5/5, AAV8, AAV25 m, AAV28 m) were compared for their transduction efficiencies using various cell lines, including BHK21, HEK293, BEAS-2BS, HeLaS3, Huh7, Hepa1-6, and A549. AAV2 and AAV1 displayed high transduction efficiency; thus, they were deemed to be suitable candidate vectors for the RI-based array. We next evaluated the impact of sodium butyrate (NaB) treatment on rAAV vector-mediated reporter gene expression and found it was significantly enhanced, suggesting that our system reflected the biological response of target cells to specific treatments.


CONCLUSIONS
Our study provides a novel method for establishing a highly efficient gene transduction array that may be developed into a platform for cell biological assays.
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Science, Fudan University, Shanghai, China.
Xiaoyan
X
Dong
Wenhong
W
Tian
Gang
G
Wang
Zheyue
Z
Dong
Wei
W
Shen
Gang
G
Zheng
Xiaobing
X
Wu
Jinglun
J
Xue
Yue
Y
Wang
Jinzhong
J
Chen
D000818:Animals; D020148:Butyric Acid / Q000494:pharmacology; D002460:Cell Line; D000229:Dependovirus / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D008854:Microscopy, Electron; D010957:Plasmids; D014161:Transduction, Genetic
D020148:Butyric Acid