System for producing synthetic promoters

Abstract

The present application discloses a method for making and selecting a transcription enhancing combined promoter cassette, which includes constructing a library of randomly combined transcription regulatory elements, which comprise double stranded DNA sequence elements that are recognized by transcription regulators; inserting the combined transcription regulatory elements upstream of a minimum promoter followed by a reporter gene in a vector; inserting the vector into a host cell; and then screening for the cells showing enhanced expression of the reporter gene, and identifying the combined promoter cassette in the cell.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] The present application is a continuation application of U.S. patent application Ser. No. 10 / 907,620, filed on Apr. 8, 2005, which claims the benefit of priority to U.S. Provisional Application No. 60 / 560,921, filed Apr. 8, 2004, the contents of which are incorporated by reference herein in its entirety.BACKGROUND OF THE INVENTION [0002] 1. Field of the Invention [0003] The present application relates to a system for designing promoters for efficient expression of genes. [0004] 2. General Background and State of the Art [0005] Despite early expectations as a promising new technology for the treatment of various diseases, convincing clinical efficacy of gene therapy has not been demonstrated in most of the trials conducted so far. One of the major technical hurdles in clinical gene therapy has been the difficulty of delivering the therapeutic gene into specific target cells and maintaining transgene expression at a sufficiently high leve...

AI Technical Summary

Benefits of technology

"The invention is about a method for efficiently producing synthetic promoters that can drive gene expression in specific cell types. This is done by creating a library of random combinations of DNA sequence elements that are recognized by transcription regulators, and then screening it using high throughput technology to select the best performing combinations. These synthetic promoters can be used in gene or virus therapy vectors to increase their efficacy and selectivity. The method can be applied to any type of cell, and it allows for the rapid introduction of new gene or virus therapy drug candidates into the market for the treatment of cancer and other diseases."

Problems solved by technology

Despite early expectations as a promising new technology for the treatment of various diseases, convincing clinical efficacy of gene therapy has not been demonstrated in most of the trials conducted so far.

One of the major technical hurdles in clinical gene therapy has been the difficulty of delivering the therapeutic gene into specific target cells and maintaining transgene expression at a sufficiently high level for a desirable amount of time.

Although targeting the delivery of therapeutic genes to specific cell types (targeted transduction) will be the most desirable, it represents a major technical obstacle and only limited success has been reported in this field (5, 24, 48).

However, these so-called cell-specific promoters are frequently leaky in terms of target cell selectivity, causing transgene expression in non-target cells at variable levels.

In most cases, these promoters are not strong enough to mediate transgene expression at or above the level required for realistic therapeutic effects.

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