A METHOD OF ENHANCED VIRAL TRANSDUCTION USING ELECTROPORATION

Abstract

Method of cell-editing comprising combining a cell or cell line with a virus, viral vector or virus like particle to form a mixture and performing simultaneous electroporation and transduction on the mixture to insert therein the virus, viral vector or virus like particle. The disclosed method simultaneously causes the virus, viral vector or virus like particle to edit, remove or modify a cell or cell line and inserting a virus, viral vector or virus like particle therein. A modified cell or cell line made by the disclosed method is also disclosed.

Specification

What is claimed is:

1. A method of enhanced viral transduction using electroporation into a cell, comprising:

■ selecting one or more cells-to-be-modified;

■ harvesting the cells-to-be-modified;

■ concentrating the cells-to-be-modified;

■ combining the cells-to-be-modified with a virus, viral vector or virus like particle to form a mixture;

■ simultaneously performing electroporation and transduction on the mixture to insert therein the virus, viral vector or virus like particle; and

■ forming one or more co-electroporated cells.

2. The method of claim 1, wherein the virus, viral vector or virus like particle is co-electroporated with gene editing agents.

3. The method of claim 2, wherein the gene editing agents are chosen from CRISPR CAS-9, RNA, plasmid, mega-TALS, gene-writing, DNase I, Benzonase, Exonuclease I, Exonuclease III, Mung Bean Nuclease, Nuclease BAL 31, RNase I, 51 Nuclease, Lambda Exonuclease, RecJ, T7 exonuclease, zinc finger nuclease, meganuclease, transcription activator-like effector nuclease, and site-specific nuclease.

4. The method of claim 1, wherein greater than 20% of the co-electroporated cells express a desired protein or peptide.

5. The method of claim 4, wherein 25-35% of the co-electroporated cells express the desired protein or peptide.

6. The method of claim 1, wherein the co-electroporated cells have a drop in viability ranging from 25-50% compared to the cells-to-be-modified.
7. The method of claim 1, wherein the co-electroporated cells have a drop in viability no more than 20% compared to the cells-to-be-modified.

8. The method of claim 1, wherein the co-electroporated cells have a drop in viability no more than 10% compared to the cells-to-be-modified.

9. The method of claim 1, wherein the co-electroporated cells have a drop in viability no more than 5% compared to the cells-to-be-modified.

10. The method of claim 1, wherein the cells-to-be-modified is within a cell population ranging from 1 x 105 to 1 x 1011.

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