Circular nucleic acid vectors, and methods for making and using the same

摘要

Circular nucleic acid vectors that provide for persistently high levels of protein expression are provided. The circular vectors of the subject invention are characterized by being devoid of expression-silencing bacterial sequences, where in many embodiments the subject vectors include a unidirectional site-specific recombination product hybrid sequence in addition to an expression cassette. Also provided are methods of using the subject vectors for introduction of a nucleic acid, e.g., an expression cassette, into a target cell, as well as preparations for use in practicing such methods. The subject methods and compositions find use in a variety of different applications, including both research and therapeutic applications. Also provided is a highly efficient and readily scalable method for producing the vectors employed in the subject methods, as well as reagents and kits/systems for practicing the same.

主权项

1. A method for providing persistent, high level expression of an expression cassette in a target cell, said method comprising:
introducing into said target cell a non-viral circular vector that is devoid of expression silencing bacterial DNA sequences and comprises said expression cassette, and testing said target cell for expression of the expression cassette more than 1 week after said introducing; wherein said vector is maintained episomally inside the target cell, and provides for persistent and high level expression of said expression cassette for greater than one week when present in said target cell.