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Published on: January 9, 2020
Cancer Cell-specific Transfection of hCas9 Gene Using Ad5F35 Vector
Wataru Matsunaga1, Katsuyuki Hamada2,3, Masatoshi Tagawa4
1Joint-use Research Facilities, Hyogo College of Medicine, Nishinomiya, Japan.
Background:
The clustered regularly interspaced short palindromic repeats-associated protein 9 (CRISPR-Cas9) is thought to have promising clinical potential. However, the off-target effects of Cas9 are a major concern for its application. Therefore, we hypothesized that the adverse effects of off-target gene editing might be minimized if the human codon-optimized Streptococcus pyogenes Cas9 (hCas9) could be specifically expressed in cancer cells.
Materials And Methods:
We constructed a chimeric adenoviral vector, Ad5F35-MKp-hCas9, and infected human bladder cancer cell lines with this vector. The confirmation of hCas9 gene expression was performed in 3-4 days after from infection.
Results:
hCas9 gene expression was observed in Ad5F35-MKp-hCas9 infected bladder cancer cells but not in non-malignant cells.
Conclusion:
Our study showed that the Ad5F35-MKp-hCas9 vector is capable of expressing the hCas9 gene with high specificity in bladder cancer cells. These findings may help in minimizing the risk of off-target effects of gene editing.
