Abstract
The attempts to attain satisfactory gene delivery ability from various engineered or non-engineered bacterial vectors have been reported by several groups. We have successfully developed a modified bactofection method for large-sized DNA delivery into human cell lines.Applying the same method of delivery, this study for the first time demonstrates an efficient and functional delivery of an 80-kb BAC clone carrying human beta-globin gene into HT1080 cell line via invasive E. coli DH10B vector. As a resultant, the modified bactofection method attained high DNA delivery even at low MOI and showed ∼5 folds increased delivery over the classic bactofection method. Upon comparing delivery efficiencies of the classic bactofection, modified bactofection, and Lipofectamine mediated transfection (10 μg/μl) at 1000 MOI, it showed 10%, 60%, and 14% GFP expression, respectively. Furthermore, functionality of the delivered BAC DNA was assessed and confirmed by RT-PCR analysis. To sum up, it can be envisioned that with careful study the modified bactofection method could successfully be employed for delivering various large-sized genes into human cells, be it for therapeutic purposes or studying their functional aspects. Besides, applying the modified bactofection method to discrete bacterial vector systems could help to achieve high efficiency DNA delivery of the large-sized genes.
| Original language | English |
|---|---|
| Article number | 103159 |
| Journal | Journal of Drug Delivery Science and Technology |
| Volume | 70 |
| DOIs | |
| State | Published - Apr 2022 |
| Externally published | Yes |
Bibliographical note
Publisher Copyright:© 2022 Elsevier B.V.
Funding
The authors acknowledge the support received from School of Sciences, Monash University , Malaysia. Alviya Sultana also acknowledge her doctoral supervisor A/P Kumaran Narayanan and co-supervisor Prof. Chow Sek Chuen, for all their support and constant guidance throughout the study. Raj Kumar is thankful to A.S. for providing an opportunity to work with her and to University of Michigan. The authors acknowledge the support received from School of Sciences, Monash University, Malaysia. Alviya Sultana also acknowledge her doctoral supervisor A/P Kumaran Narayanan and co-supervisor Prof. Chow Sek Chuen, for all their support and constant guidance throughout the study. Raj Kumar is thankful to A.S. for providing an opportunity to work with her and to University of Michigan.
| Funders | Funder number |
|---|---|
| School of Sciences | |
| University of Michigan | |
| Monash University Malaysia |
Keywords
- BAC DNA
- Beta-globin
- GFP expression
- Modified bactofection
- gene delivery
- gene therapy