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CSIC BIO029- Method for efficient insertion at specific sites into any target DNA of group II introns
Zaidin Experimental Agricultural Station of CSIC has identified a group II intron for inserting a nucleic acid molecule into any target DNA. This technology has been filled as Spanish Patent, but we are into the priority year. The main advantage is that these intron sequences can be modified in order to redirect it to a chosen target DNA site and could be used for mutagenesis, controlling gene expression, gene tragging, etc. The Research Team are looking for a company interested in obtaining a licence agreement and invest in the development of specific uses.
Specification sheet:
DESCRIPTION

The invention employs a group II intron for inserting a nucleic acid molecule into any target DNA. The DNA target site for these type of introns requires a DNA region encompassing 25 nucleotides in which 14 nt are recognised primarily by base pairing with sequences contained in two loops of the the intron RNA (EBS sequences). These intron sequences can be modified in order to redirect it to a chosen target DNA site.

Group II introns are catalytic RNAs that function as mobile genetic elements by inserting directly into specific DNA target sites. This mobility is mediated by the RNA component plus a multifunctional intron-encoded protein (IEP) that has reverse transcriptase (RT), RNA splicing (maturase) and in some mobile introns, a DNA endonuclease activity. In addition, the bacterial group II introns mobility appears to be independent of recombinase. Thus, group II introns are good candidates for genetic engineering applications of great importance in living organisms with deficient recombination pathways like eukaryotes.

The DNA target site for these type of introns requires a DNA region encompassing 25 nucleotides in which 14 nt are recognised primarily by base pairing with sequences contained in two loops of the the intron RNA (EBS sequences). These sequences can be modified in order to redirect this intron to a chosen target DNA site.

The present invention provides new methods, employing a group II intron for inserting a nucleic acid molecule into any target DNA at specific sites. The methods comprise the step of: providing a group II intron RNA having two hybridizing sequences that are capable of hybridizing with two intron RNA binding sequences on the one strand of the DNA substrate, modifying the group II intron RNA hybridizing sequences to hybridizes with specific sequences of one strand in any target DNA, and providing an intron-encoded protein, which naturally lacks the endonuclease domain.

Among applications retargeted group II introns can be used for: mutagenesis, controlling gene expression, gene tagging, repairing a non-functional gene and for biological containment.

INNOVATIVE ASPECTS

The present invention provides a new strategy for modifying genes employing the `homing` (mobile) ability of group II introns (in some cases 100% homing frecuency). This ability together with the DNA target specificity of this mobile elements makes them an excellent alternative to the mutagenic systems described so far (mainly transposons, ...) with potential applications in genetic engineering and functional genomics.

COMPETITIVE ADVANTAGES

1) A higher mobility efficiency (3 or 4 exponential orders) than the described for transposons.

2) Specificity of the target. A few genomic locations correspond to 14 nt specific sequence.

3) The mobility occurs independently of homologous recombination systems.

KEYWORDS

Mutagenic-system, functional-genomics, Group II introns, Retroelements, Genetic-engineering, gene therapy.

PATENT

Applied for

CONTACT

Consuelo Quilchano
email: quilchano@cica.es
phone: +34 95 450 09 78

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