Hi, I am currently working on CAR cell therapeutics development and I am using lentivirus as a vector for transgene delivery.
As per my understanding, lentiviral transduction is known to give persistent expression of GOI compared to transient transfection such as electroporation.
However, when I deliver CAR gene, expression level of CAR is decreased over time and nearly no expression is detected after one week and same result were observed on several trials.
If I may, could you kindly let me know the methods that I can conduct to increase the persistency of my CAR expression level for a longer period of time?
The persistence of lentiviral vector expression relies on active viral replication or integration into the genome. If integrated into the genome the viral sequences will encourage gene silencing through methylation. It is therefore recommended to add insulator sequences either side of your GOI. If you want persistence as an extrachromosomal element then you will need to add an active origin of replication (or two ssos) and DTS or artificial centrosome. A cyclovirus based origin might be beneficial as you can use two origins flanking the replicase and your GOI to make a miniplasmid in the destination cells that has lost the lentiviral backbone.
- Badges
- Science topic
- Similar topics
- Biological Science
- Immunology
- Antibodies