RIKEN DNA Bank Adenovirus
Living modified organisms (LMO)
Laboratory use only.
Not intended for intentional introduction into the environment.
Transportation of recombinant adenovirus
For US transfer, "Veterinary Permission" (VS Forms 16-3 and 16-7) might be required for shipment of the biological materials. Please refer to our website at http://dna.brc.riken.jp/en/furnish.html.
RIKEN DNA Bank provides recombinant adenoviruses as crude lysates of infected cells with 106-8 pfu/ml virus particle. The volume of lysate provided is 0.1 ml.
Based on results of our research (Ugai et al., 2002), recombinant adenoviruses are provided at room temperature by air mail.
After received a sample, amplify the adenovirus immediately by infection of 9x106 cells of HEK293 in 10cm diameter dish with the 100 ul virus solution. Otherwise you save it at -80oC.
Refer a protocol for purification or our published paper (Ugai et al., 2005).
Replication Competent Adenovirus (RCA)
Recombinant adenoviruses provided by RIKEN DNA Bank are prepared by the 'COS-TPC method' (Miyake et al., 1996) and, in some cases, they might be suspected of contamination by Replication Competent Adenovirus (RCA) even though they were inspected by the "RCA-checking" test (Suzuki et al., 2004) before shipment. In addition, it was reported that RCAs harboring the E1 gene (E1A and/or E1B) derived from HEK293 cells were found in solutions of virus during the preparation of high-titer stocks (Ugai et al., 2003). For these reasons, tests for contamination by RCAs should be performed prior to use of recombinant adenoviruses.
Moreover, recombinant adenoviruses, as well as recombinant DNA clones, can potentially mutate in some cases. Inserted genes (and the fiber gene, in the case of fiber-substituted recombinant adenoviruses) should be examined prior to your experiments by PCR, nucleotide sequencing and/or Western blotting.
You can find recombinant adenoviruses at
Entire list of viruses and shuttle vectors are avairable.
Link & References
- Introduction of Recombinant Virus Bank in RIKEN Gene EngineeringDivision by Takehide Murata, Jianzhi Pan, Megumi Hirose, KumikoInabe, Yukari Kujime, Chitose Kurihara, Yuka Kusa, Satoko Masuzaki,Koji Nakade, Yuri Nakano, Masato Ohkubo, Takahito Yamasaki, YuichiObata, Kazunari K. Yokoyama. Published in the Gene Therapy Review.
- Fukuda, H., Terashima, M., Koshikawa, M., Kanegae, Y., Saito, I. (2006) Possible mechanism of adenovirus generation from a cloned viral genome tagged with nucleotides at its ends.
Microbiol Immunol. 50: 643-654.
- Miyake, S., Makimura, M., Kanegae, Y., Harada, S., Sato, Y., Takamori, K., Tokuda, C., Saito, I. (1996) Efficient generation of recombinant adenoviruses using adenovirus DNA-terminal protein complex and a cosmid bearing the full-length virus genome.
Proc. Natl. Acad. Sci. USA 93: 1320-1324.
- Suzuki, E., Murata, T., Watanabe, S., Kujime, Y., Hirose, M., Pan, J., Yamazaki, T., Ugai, H., Yokoyama, K.K. (2004) A simple method for the simultaneous detection of E1A and E1B in adenovirus stocks.
Oncol. Rep. 11: 173-178.
- Ugai, H., Watanabe, S., Suzuki, E., Tsutsui-Nakata, H., Yokoyama, K.K., Murata, T. (2002) Stability of a recombinant adenoviral vector: optimization of conditions for storage, transport and delivery.
Jpn. J. Cancer Res. 93: 598-603.
- Ugai, H., Suzuki, E., Inabe, K., Murata, T., Hamada, H., Yokoyama, K.K. (2003) Spontaneous mutations in the human gene for p53 in recombinant adenovirus during multiple passages in human embryonic kidney 293 cells.
Biochem. Biophys. Res. Commun. 300: 448-456.
- Ugai H, Yamasaki T, Hirose M, Inabe K, Kujime Y, Terashima M, Liu B, Tang H, Zhao M, Murata T, Kimura M, Pan J, Obata Y, Hamada H, Yokoyama KK.(2005) Purification of infectious adenovirus in two hours by ultracentrifugation and tangential flow filtration.
Biochem Biophys Res Commun. 331: 1053-1060.
- Zhu, J., Grace, M., Casale, J., Chang, A.T., Musco, M.L., Bordens, R., Greenberg, R., Schaefer, E., Indelicato, S.R. (1999) Characterization of replication-competent adenovirus isolates from large-scale production of a recombinant adenoviral vector.
Human Gene Ther. 10: 113-121.
Efficiency of infection of adenovirus
- Wickham et al., Nat. Biothech. 14, 1570-1573, 1996.
- Mizuguchi and Hayakawa, Gene 285, 69-77, 2002.
- Li et al., Mol. Cancer Ther. 4, 1850-1859, 2005.
- Huch et al., Hum. Gene Ther. 17, 1-14, 2006.
- Yoshida et al., Hum. Gene Ther. 9, 2503-2515, 1998.
- Nakamura et al., Hum. Gene Ther. 13, 613-626, 2002.
- Steinwaerder et al., Hum. Gene Ther. 11, 1933-1948, 2000.
- Dehari et al., Cancer Gene Ther. 10, 75-85, 2003.
- Bruning and Runnebaum, Gene Ther. 10, 198-205, 2003.
- Shayakhmetov and Lieber, J. Virol. Nov, 10274-10286, 2000.
- Israel et al., J. Virol. June, 5215-5221, 2001.
- Michiels F et al., Nat. Biotechnol. 20, 1154-1157, 2002.
- Michiels F et al. (2002). Arrayed adenoviral expression libraries for functional screening. Nat. Biotechnol. 20: 1154-1157.
|Table of infection efficiency of human cells and cell lines with standard Ad 5 viruses at the indicated Multiplicity of Infection (MOI). (form Michiels F et al. (2002), Nat. Biotechnol. 20: 1154-1157)
||Origin of cells
||Human liposuction tissue
|Bronchial epithelial cells
||Cultured from human CD34+ cord blood cells
||Human peripheral blood
||Human peripheral blood, naive
||Differentiated mesenchymal stem cells
||Human umbilical cord vascular endothelium
||Human pluripotent bone marrow progenitors
||Human liposuction tissue
||Human foreskin fibroblasts
||Human lung carcinoma
||Human cervix tumor
||Human liver carcinoma
||Human T lymphoma
||Human erythroid leukemia
||Human breast cancer
||Human B-cell lymphoma
||Human neuronal tissue
||Human breast cancer