Crowding agent-induced nucleic acid transfer into a recipient host cell
AssigneeSynthetic Genomics, Inc.
InventorsBogumil J. Karas, Hutchison Clyde A., III, Hamilton O. Smith, Yo Suzuki
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Abstract
The presently disclosed invention relates to methods of transferring large nucleic acid molecules or a genome from one cell (the donor) into heterologous host cells in the presence of a crowding agent. The method allows for greater ease and efficiency of transfer of genetic material. Introduction of the donor genetic material into the recipient host cells also allows for manipulation of the donor nucleic acid molecule or genome within the host cells. Methods disclosed herein can be used to alter donor genomes from intractable donor cells in more tractable host cells.
Claims26
A method for donor cell nucleic acid or genome installation in heterologous host cells comprising:
- obtaining a population of donor cells comprising said donor nucleic acid or genome, wherein said donor nucleic acid or genome is an essentially intact nucleic acid that is at least a minimal genome, and is greater than about 150 kb in length;
- and
- contacting said population of donor cells with a population of heterologous host cells in the presence of from about 12% (w/w) to about 20% (w/w) of a crowding agent, thereby generating a population of host cells comprising the donor nucleic acid or genome.
A method for donor cell nucleic acid or genome installation in heterologous host cells comprising:
- (a) obtaining a population of donor cells comprising said donor nucleic acid or genome, wherein said donor nucleic acid or genome is an essentially intact nucleic acid that is at least a minimal genome, and is greater than about 150 kb in length;
- and
- (b) contacting said population of donor cells with said population of heterologous host cells in the presence of about 15% or more of a crowding agent, thereby generating a population of host cells comprising the donor genome;
- wherein step (c) does not include encapsulation of the donor nucleic acid or genome in an agarose plug prior to contacting said donor cells with said population of heterologous host cells.
Dependencies are inferred from the claim text.