Methods for sustained and regulatable gene expression using viral based expression vectors

摘要

Disclosed are genetic expression cassettes, and vectors comprising them useful for the delivery of isolated nucleic acid segments including those expressing or encoding one or more selected therapeutic constructs (including, without limitation, therapeutic peptides, polypeptides, ribozymes, or catalytic RNA molecules), to one or more selected cells or tissues of a vertebrate animal. Methods employing the disclosed genetic constructs in the development of gene therapy-based viral vector systems are also disclosed. The expression cassettes and viral vectors disclosed herein provide new tools for methods of treating mammalian, and in particular, human diseases, disorders, and/or dysfunctions. The disclosed compositions and methods find particular utility in a variety of investigative, diagnostic, and therapeutic regimens, including, for example, in the treatment or amelioration of symptoms of a variety of mammalian, and particularly, human conditions.

主权项

1. A method for expressing a heterologous nucleic acid segment of interest in a mammalian host cell, comprising providing to a population of mammalian cells, an expression vector comprising, in 5′ to 3′ order,
(a) a first isolated HSV LAT insulator/boundary region that consists essentially of a contiguous nucleotide sequence from about nucleotide 8365 to about nucleotide 9273 of SEQ ID NO:102, SEQ ID NO:103, or SEQ ID NO:104, operably positioned 5′ of a first polynucleotide comprising the heterologous nucleic acid segment of interest operably linked to a first promoter that expresses the heterologous nucleic acid segment in one or more cells of the population; and (b) a second isolated HSV LAT insulator/boundary region that consists essentially of a contiguous nucleotide sequence from about nucleotide 120,208 to about nucleotide 120,940 of SEQ ID NO:102, SEQ ID NO:103, or SEQ ID NO:104, operably positioned 3′ of the heterologous nucleic acid segment of interest; in an amount and for a time effective to express the heterologous nucleic acid segment in one or more cells of the population.