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electroporation transfer

Electroporation transfer is emerged as a powerful tool for the genetic modification of diverse cell types based on the transient disruption of cell membrane via exposure to an electric field, which allows charged molecules to enter the cell. For instance, the square-wave pulse-based new electroporation devices, such as Lonza Nucleofector II electroporation system, manifests a high efficiency in the genetic modification of T cells with proprietary electroporation buffers and electric parameters. https://www.creative-biolabs.com/car-t/gene-packaging-delivery.htm

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electroporation transfer

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  1. CAR-T Gene Packaging & Delivery Reference: https://www.creative-biolabs.com/car-t/gene-packaging-delivery.htm

  2. Background • Since the successful manufacture of CAR-T cells is largely dependent on T cell receptor (TCR) gene transfer, this genetic modification of T lymphocytes is one of the most critical steps to generate superior CAR-T cells. Although the T cells are difficult to modify using non-viral methods like lipid-based car t plasmid transfection due to the high toxicity and low efficiency, the electroporation transfection and transposon based transfection systems are much more robust to complete the T-cell transfection.

  3. CAR-T cell transfection approaches

  4. Electroporation • Electroporation is emerged as a powerful tool for the genetic modification of diverse cell types based on the transient disruption of cell membrane via exposure to an electric field, which allows charged molecules to enter the cell. • For instance, the square-wave pulse-based new electroporation devices, such as Lonza Nucleofector II electroporation system, manifests a high efficiency in the genetic modification of T cells with proprietary electroporation buffers and electric parameters. In detail, up to 80 % of viability and 40-60 % of expression are achieved in human T cells.

  5. Retroviral transfection • one of the mainstays of current gene therapy approaches, which contains a reverse transcriptase to enable the integration of artificial genes into the host genome in a stable status. Particularly, the retroviral vectors can either be replication-competent or replication-defective, and the latter is the most common choice for research since the viruses are competent for additional rounds of virion replication and packaging with other genes. • With a typical maximum 8-10 kB length of an allowable DNA insertion in a replication-defective viral vector, the viruses are capable of infecting the target cells and delivering the specific viral payload. Meanwhile, the MLV fails to perform the typical lytic pathway and lead to cell lysis and death. Noticeably, MLV requires actively dividing cells for transduction.

  6. Lentiviral (complex retrovirus) transfection • one of the mainstays of current gene therapy approaches, which enable the integration of artificial genes into the host genome in a stable status for genetical modifications in actively dividing cells as a subclass of retroviruses. • More powerfully, the lentivirus has the ability to integrate foreign genes into the genome of non-dividing cells. Generally comparing to the electroporation, the virus-based transfections are relatively time-consuming and expensive, which limit a broader application in the clinical usage.

  7. Thanks Reference: https://www.creative-biolabs.com/car-t/gene-packaging-delivery.htm

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