Life Technologies Corporation

Invitrogen Corporation

Valentina C Ciccarone

Alberto Haces

Joel A Jessee

Pamela Hawley-Nelson

Gulilat Gebeyehu

Jianqing Lan

Dale F Gruber

PoJen Shih

Kevin P Schifferli

Krista L Evans

Shelly Bennett

Sharon Cates

Anna Chytil

The present invention is directed generally to cell culture media useful for introducing macromolecules and compounds (e.g., nucleic acid molecules) into cells (e.g., eukaryotic cells) in the presence of said media. Cells containing introduced materials can be further cultured in the media. In particular, the invention allows introduction of nucleic acid molecules (e.g., vectors) into cells (particularly eukaryotic cells) and expression of proteins encoded by the nucleic acid molecules in the cells. The invention obviates the need to change the cell culture medium each time a different procedure is performed with the cells (e.g., culturing cells vs. transfecting cells). The invention thus provides efficient and high throughput methods to transform/transfect culture and cells avoiding the need for multiple manipulations and transfers of cells during transfection and expression studies.

Culture medium for cell growth and transfection

The present invention provides compositions useful for transfecting eukaryotic cells comprising nucleic acid complexes with peptides, wherein the peptide is optionally covalently coupled to a nucleic acid-binding group, and cationic lipids or dendrimers as transfection agents. The invention also provides transfection compositions in which a peptide is covalently linked to the transfection agent (lipid, cationic lipid or dendrimer). Inclusion of peptides or modified-peptides in transfection compositions or covalent attachment of peptides to transfection agents results in enhanced transfection efficiency. Methods for the preparation of transfection compositions and methods of using these transfection compositions as intracellular delivery agents and extracellular targeting agents are also disclosed.

Peptide-enhanced transfections

The present invention discloses highly packed polycationic ammonium, sulfonium and phosphonium lipid compounds useful for making lipid aggregates for delivery of macromolecules and other compounds into cells. They are especially useful for the DNA-dependent transformation of cells. Methods for their preparation and use as intracellular delivery agents are also disclosed.

Highly-packed polycationic ammonium, sulfonium and phosphonium lipids

The present invention provides novel cell lines that may have improved adhesive qualities, transgene expression level, growth rate, and/or growth rate in serum free medium or even chemically defined compared to cells of the prior art.

Attached cell lines

Highly packed polycationic ammonium, sulfonium and phosphonium lipids

The present invention describes cells and constructs for a regulated viral (e.g. alphavirus) expression system, where gene expression is controlled by controlling expression of replicases or nonstructural proteins and/or controlling the amount of such proteins introduced in a cell, which in turn regulates RNA replication and subsequently gene expression. Particularly, this system takes advantage of the high level expression of the alphavirus systems for recombinant protein production and allows for large scale applications without biosafety concerns.

Regulated expression of recombinant proteins using RNA viruses

The present invention discloses cationic lipids useful for making lipid aggregates for delivery of macromolecules and other compounds into cells. They are especially useful for the DNA-dependent transformation of cells. Also disclosed are lipids useful both for the delivery of macromolecules and also useful as intermediates for making other such lipids.

Growing community of inventors

Gaithersburg, MD, United States of America

Valentina C Ciccarone