Efficient Large Volume Lentiviral Vector Production Using Flow Electroporation

Witting, Scott R.; Li, Lin Hong; Jasti, Aparna C.; Allen, Cornell; Cornetta, Kenneth; Brady, John Paul; Shivakumar, Rama; Peshwa, Madhusudan V.

doi:10.1089/hum.2011.088

Cited by 32 publications

(22 citation statements)

References 45 publications

(53 reference statements)

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“…To demonstrate that NanoMEDIC production is scalable and adaptable for xeno-free conditions for future clinical applications, we developed a HEK293T suspension cell production system utilizing flow electroporation by the MaxCyte STX, previously reported for large-scale lentivirus production 39 .…”

Section: Nanomedic Efficiently Edits Various Cells To Show Thatmentioning

confidence: 99%

Extracellular nanovesicles for packaging of CRISPR-Cas9 protein and sgRNA to induce therapeutic exon skipping

et al. 2020

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Prolonged expression of the CRISPR-Cas9 nuclease and gRNA from viral vectors may cause off-target mutagenesis and immunogenicity. Thus, a transient delivery system is needed for therapeutic genome editing applications. Here, we develop an extracellular nanovesicle-based ribonucleoprotein delivery system named NanoMEDIC by utilizing two distinct homing mechanisms. Chemical induced dimerization recruits Cas9 protein into extracellular nanovesicles, and then a viral RNA packaging signal and two self-cleaving riboswitches tether and release sgRNA into nanovesicles. We demonstrate efficient genome editing in various hardto-transfect cell types, including human induced pluripotent stem (iPS) cells, neurons, and myoblasts. NanoMEDIC also achieves over 90% exon skipping efficiencies in skeletal muscle cells derived from Duchenne muscular dystrophy (DMD) patient iPS cells. Finally, single intramuscular injection of NanoMEDIC induces permanent genomic exon skipping in a luciferase reporter mouse and in mdx mice, indicating its utility for in vivo genome editing therapy of DMD and beyond.

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Section: Nanomedic Efficiently Edits Various Cells To Show Thatmentioning

confidence: 99%

Extracellular nanovesicles for packaging of CRISPR-Cas9 protein and sgRNA to induce therapeutic exon skipping

et al. 2020

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“…Another cost-effective compound, polyethylenimine, has also been qualified and used in recent years 91,92 as well as flow electroporation. 93 Other lipidbased methods are still too expensive to be used in a large-scale manufacturing setting. For large-scale lentiviral vector production, HEK293-derived cells are expanded in large quantity.…”

Section: Expression Vectors For Genetic Modification Of T Cellsmentioning

confidence: 99%

Manufacture of tumor- and virus-specific T lymphocytes for adoptive cell therapies

Wang

Rivière

2015

Cancer Gene Ther

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Adoptive transfer of tumor-infiltrating lymphocytes (TILs) and genetically engineered T lymphocytes expressing chimeric antigen receptors (CARs) or conventional alpha/beta T-cell receptors (TCRs), collectively termed adoptive cell therapy (ACT), is an emerging novel strategy to treat cancer patients. Application of ACT has been constrained by the ability to isolate and expand functional tumor-reactive T cells. The transition of ACT from a promising experimental regimen to an established standard of care treatment relies largely on the establishment of safe, efficient, robust and cost-effective cell manufacturing protocols. The manufacture of cellular products under current good manufacturing practices (cGMPs) has a critical role in the process. Herein, we review current manufacturing methods for the large-scale production of clinical-grade TILs, virus-specific and genetically modified CAR or TCR transduced T cells in the context of phase I/II clinical trials as well as the regulatory pathway to get these complex personalized cellular products to the clinic.

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“…The transfection is performed either by the calcium-phosphate precipitation method or by using PEI [95]. Rather recently, suspension based transfection processes have been evaluated using either PEI based transfection [96] or flow electroporation which has the advantage of performing transfection without the concomitant addition of transfection agents [97]. Large-scale production runs are often harvested one-to three-times while maintaining the overall vector productivity, whereas for research grade productions up to five harvests may be performed.…”

Section: Production Of Lentiviral Vectors Based On the Use Of Transiementioning

confidence: 99%

Manufacturing of viral vectors for gene therapy: part I. Upstream processing

Merten¹,

Schweizer²,

Chahal³

et al. 2014

Pharmaceutical Bioprocessing

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Efficient Large Volume Lentiviral Vector Production Using Flow Electroporation

Cited by 32 publications

References 45 publications

Extracellular nanovesicles for packaging of CRISPR-Cas9 protein and sgRNA to induce therapeutic exon skipping

Extracellular nanovesicles for packaging of CRISPR-Cas9 protein and sgRNA to induce therapeutic exon skipping

Manufacture of tumor- and virus-specific T lymphocytes for adoptive cell therapies

Manufacturing of viral vectors for gene therapy: part I. Upstream processing

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