Introduction

The invention describes a nano-transducer able to modify a target genomic sequence in a cell, in order to eliminate harmful mutations underlying genetic diseases. The new Nano-gene genomic editing tool overcomes the limitations of CRISPR/Cas9 systems by replacing the nuclease activity of Cas9 with a nano-transducer. The nano-transducer activation is bound by the occurrence of a series of conditions, giving the system greater reliability and specificity.

Technical features

The main innovative aspect of Nano-gene is that the nano-transducer consists of a hybrid system, which includes: 1) a nanoparticle covalently bound to a g-RNA/dCas9 (a mutated Cas9 able to bind DNA to the target site via g-RNA but unable to cut DNA) and 2) a restriction enzyme that only activates at 65 °C. The nanoparticle is designed to absorb energy at a specific wavelength and convert it into heat. Activation of the nano-transducer, as a result of  laser irradiation, causes localized temperature increase, only in the gene locus recognized by ribonucleoprotein. At 65 °C the activated restriction enzyme causes the double-stranded DNA to be cut into the chosen gene locus. The supremacy of Nano-gene technology over current methodologies lies in multi-function integration: time (editing occurs only when the laser is on), space (editing occurs only where the laser is focused) and the fidelity function (editing only takes place if the nano-transducer is on-target).

Possible Applications

• Medicine;​
• Gene therapy;​
• Treatment of pathologies triggered by specific DNA aberrations:​ cancer, rare diseases.

Advantages

• High specificity on the target gene;​
• Minimization of side effects, as non-specific genome cutting;​
• Independence from the activity of Cas9;​
• Limited activity of Cas9 on genome exposure;​
• Reduced toxicity compared to CRISPR/Cas9; ​
• Compatibility with a multi-gene treatment.

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