Regeneron Pharmaceuticals, Inc.

Regeneran Pharmaceuticals, Inc.

Gregg S Warshaw

Jose F Rojas

Andrew J Murphy

Lynn Macdonald

David M Valenzuela

John R McWhirter

Ka-Man Venus Lai

Corey Momont

Chris Schoenherr

Caitlin Montagna

Chia-Jen Siao

Susannah Brydges

Caitlin L Goshert

John Mcwhirter

Methods are provided herein for assembling at least two nucleic acids using a sequence specific nuclease agent (e.g., a gRNA-Cas complex) to create end sequences having complementarity and subsequently assembling the overlapping complementary sequences. The nuclease agent (e.g., a gRNA-Cas complex) can create double strand breaks in dsDNA in order to create overlapping end sequences or can create nicks on each strand to produce complementary overhanging end sequences. Assembly using the method described herein can assemble any nucleic acids having overlapping sequences or can use a joiner oligo to assemble sequences without complementary ends.

Nuclease-mediated DNA assembly

Methods for introducing a scarless targeted genetic modification into a preexisting targeting vector are provided. The methods can use combinations of bacterial homologous recombination (BHR) and in vitro assembly to introduce such targeted genetic modifications into a preexisting targeting vector in a scarless manner.

Methods for scarless introduction of targeted modifications into targeting vectors

Growing community of inventors

Bayside, NY, United States of America

Gregg S Warshaw