A rapid, efficient, and cost-effective method for titering third-generation lentiviral vectors

Abstract

Lentiviral vectors are useful vectors for stable transduction and permanent expression in dividing and non-dividing cells. In particular, third-generation lentiviral vectors have been engineered to be significantly safer than their second-generation counterparts, incorporating several safety features not present in earlier versions. For example, the tat gene, which is essential for the replication of wild-type human immunodeficiency virus type 1, has been deleted, and vector packaging functions have been distributed across three separate plasmids, further enhancing safety. In both research and clinical settings, having a reliable and accurate method for titering lentiviral vectors is critical. We have developed a method using the Woodchuck Hepatitis Virus Post-transcriptional Regulatory Element as a template for a real-time quantitative polymerase chain reaction, coupled with TRIzol lysis buffer for ribonucleic acid isolation. This method yielded results comparable to those from a commonly used commercial kit, offering advantages of speed, cost-effectiveness, and accuracy. It presents a viable, economical alternative for both research and clinical laboratories.

Keywords

Titer / Third-generation lentiviral vector / Molecular biology / Ribonucleic acid / TRIzol

Funding

This work was supported by the Ideas Grant (no. 1187040), funded by the National Health and Medical Research Council of Australia.

Conflict of interest

The authors declare that they have no competing interests.

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