Generation Bio Co.

Matthew G Stanton

Birte Nolting

Brian W Budzik

Ozan Alkan

Douglas Anthony Kerr

Robert Michael Kotin

Carolyn Pelletier

Andrew Milstead

Hongbiao Liao

Gregory L Beutner

The application describes methods for synthetic synthesis and cell-free synthesis of DNA vectors, particularly closed-ended DNA vectors (e.g., ceDNA vectors) having linear and continuous structure for delivery and expression of a transgene. The present invention relates to an in vitro process for production of closed-ended DNA vectors, corresponding DNA vector products produced by the methods and uses thereof, and oligonucleotides and kits useful in the process of the invention. DNA vectors produced using the methods described herein are free from unwanted side effects due to contaminants introduced during production in cell lines, for example, bacterial or insect cell lines. Further provided herein are methods and cell lines for reliable gene expression in vitro, ex vivo and in vivo using the ceDNA vectors synthesized using the methods herein.

Closed-ended DNA vectors obtainable from cell-free synthesis and process for obtaining ceDNA vectors

Provided herein are lipids having the Formula I or Formula Ia: and pharmaceutically acceptable salts thereof, wherein R', R, R, R, R, R, R, R, X, X, and n are as defined herein for Formula I and Formula Ia, respectively. Also provided herein are lipid nanoparticle (LNP) compositions comprising lipid having the Formula I or Ia and a capsid-free, non-viral vector (e.g., ceDNA). In one aspect of any of the aspects or embodiments herein, these LNPs can be used to deliver a capsid-free, non-viral DNA vector to a target site of interest (e.g., cell, tissue, organ, and the like).

Lipids and compositions thereof

Provided herein are lipids having the Formula (I): and pharmaceutically acceptable salts thereof, wherein R, R, a, and b are as defined herein. Also provided herein are lipid nanoparticle (LNP) compositions comprising lipid having the Formula (I) and a capsid-free, non-viral vector (e.g., ceDNA). In one aspect of any of the aspects or embodiments herein, these LNPs can be used to deliver a capsid-free, non-viral DNA vector to a target site of interest (e.g., cell, tissue, organ, and the like).

Lipids and nanoparticle compositions thereof

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Cambridge, MA, United States of America

Matthew G Stanton