Construction of Lentiviral Vectors for CAR-T Cell Applications in Mongolia

DOI

10.2991/978-94-6463-837-0_14How to use a DOI?

Keywords

Cancer Immunotherapy; Genetic Engineering; Vector Construction; Plasmid; Transfection

Abstract

CAR-T cell therapy is a cutting-edge form of immunotherapy that involves extracting T cells from a patient, genetically modifying them into CART cells, and then reinfusing them to the target and to eliminate cancer cells. Lentiviruses are commonly used in this process to deliver the chimeric antigen receptor (CAR) gene into T cells, enabling them to recognize and attack cancer cells. Recombinant lentiviral vectors are especially advantageous because they can transduce both dividing and non-dividing cells, are generally considered safe, and provide long-term transgene expression. This is due to the integration of the viral genome known as the provirus into the host cell’s DNA, allowing it to be passed on to daughter cells. These properties are particularly important for CAR-T cell therapy, where stable and sustained expression of the CAR is essential. Lentiviral vectors are often used to deliver protein-encoding cDNAs, such as CAR constructs or reporter genes, as well as noncoding sequences for RNA interference or genome editing, including shRNA or gRNA. This study aimed to construct and validate lentiviral vectors for use in CAR-T cell therapy. Four essential plasmids—pMDLg/pRRE, pRSV-Rev, pCMV-VSV-G, and a CAR-encoding plasmid—were successfully amplified using competent cells. The plasmids were then purified, quantified, and transfected into HEK293T cells using PEI STAR. Lentiviral particles were harvested post-transfection and confirmed by RT-qPCR. In HEK293T cells, transduction efficiency reached 24.5%, while transfection efficiency was measured at 80%, as determined by fluorescence microscopy and flow cytometry. These results confirm the successful construction of functional lentiviral vectors suitable for downstream CAR-T cell applications. This establishes a foundational platform for CAR-T cell applications development in Mongolia.

Copyright

© 2025 The Author(s)

Open Access

Open Access This chapter is licensed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/), which permits any noncommercial use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license and indicate if changes were made.

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TY  - CONF
AU  - Suvd-Erdene Uurdmunkh
AU  - Gansukh Choijilsuren
AU  - Ulziikhuu Tumurkhuyag
AU  - Ariunzaya Bat-Erdene
AU  - Ulziisaikhan Bat-Itgel
AU  - Urangoo Bayasgalanbaatar
AU  - Batbaatar Gunchin
AU  - Enkhsaikhan Lkhagvasuren
AU  - Tsogtsaikhan Sandag
PY  - 2025
DA  - 2025/09/17
TI  - Construction of Lentiviral Vectors for CAR-T Cell Applications in Mongolia
BT  - Proceedings of the International Biological Conference in Mongolia 2025 (IBCM 2025)
PB  - Atlantis Press
SP  - 199
EP  - 210
SN  - 2468-5747
UR  - https://doi.org/10.2991/978-94-6463-837-0_14
DO  - 10.2991/978-94-6463-837-0_14
ID  - Uurdmunkh2025
ER  -