Nucleic acids, compositions and methods for the excision of target nucleic acids

Abstract

Nucleic acids, compositions, and methods that allow for the excision of one or more loci from the genome of a host cell are provided herein. In particular, provided herein is an excisable nucleic acid construct comprising, in a 5' to 3' orientation: a first tandem repeat nucleic acid, a first homing endonuclease recognition site, a target nucleic acid, a second homing endonuclease recognition site, and a second tandem repeat nucleic acid. In some embodiments, the excisable nucleic acid construct is integrated into the host cell genome, and the target nucleic acid can be excised from the host cell genome by contacting the homing endonuclease recognition sites with one or more appropriate homing endonucleases.

Description

[0001] This application claims the priority of U.S. Provisional Application No. 61 / 378,350 whose filing date is August 30, 2010, the entire content of which is incorporated into this application by reference. 1. Technical Field [0002] The nucleic acids, compositions, and methods provided in this application generally involve the fields of molecular biology and genetic engineering. 2. Background technology [0003] In many fields, including metabolic engineering, industrial microbiology, synthetic biology, and basic molecular genetics research, it is necessary to use genetic engineering techniques to excise target nucleic acids from the host cell genome or episomal genes. However, the methods previously used to remove target nucleic acids have been restricted and limited in application. For example, the use of site-specific recombinase methods to remove harmful specific recombinase binding sites will leave behind, and these sites will create potential genomic instability in the h...

AI Technical Summary

Problems solved by technology

Other methods produce excision events at a lower frequency, so it is necessary to find ways to select for rare host cells undergoing excision events by growth

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