Regulated genetic suicide mechanism compositions and methods

摘要

Embodiments of the present invention relates to the incorporation and use of a regulated genetic suicide mechanism for use in the improved purification of biologics, including adjunct use in various eubacterial minicell production and purification methodologies. Described herein are high-yield eubacterial minicell-producing strains with genetic modifications that comprise a regulated genetic suicide mechanism that irreparably destroys the parent cell chromosome such that live parental cells in a culture can be functionally eliminated at any time during the course of a minicell production and purification run. Embodiments of the present invention also describe methods useful in the elimination of live parental cells during the production of other cell-based biologics.

主权项

1. A eubacterial minicell derived from a minicell-producing bacteria, wherein the minicell-producing bacteria comprises:
an expressible cell division gene, wherein the cell division gene is selected from the group consisting of zipA, sulA, secA, dicA, dicB, dicC, dicF, ftsA, ftsI, ftsN, ftsK, ftsL, ftsQ, ftsW, ftsZ, minC, minD, minE, seqA, ccdB, sfiC, and ddlB; and an expressible gene encoding a homing endonuclease, wherein the homing endonuclease gene is a transgene, and wherein the chromosome of the minicell-producing bacteria comprises one or more recognition sites of the homing endonuclease.