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I-Gene tools for gene ] [ editing

The I·Gene technology and the following I·Geneer Kit offers a rationally designed approach for highly specific and precise genome editing tool at the in vitro level. It presents radically new concept of gene editing, in which the nuclease activity of Cas9 is replaced by a so called nanotransducer (NT), whose activation is constrained by a number of conditions being realized by the following elements:

  • The DNA recognition element (sensor)- a module responsible to guide the NT in the desired genomic location performed by catalytically-dead Cas9 (dCas9) [3]. The D10A/H840A point mutation abolishes Cas9 nuclease activity (dCas9 cannot cut DNA) but does not disable the binding activity of Cas9 to the g-RNA and RNA-guided DNA unwinding activity (dCas9 can recognise the target DNA sequence).

  • The nanotransducer (NT) - a plasmonic nanoparticle covalently linked to dCas9/g-RNA complex, capable of absorbing a specific optical wavelength and efficiently converting it into nano range heat radiation.

  • The activator - a heat radiation generated by the NT upon laser impulse (or any other controlled temperature increase), which is used to faciliate DNA cleavage.



[1] Chira S, Gulei D, et al. CRISPR/Cas9: Transcending the Reality of Genome Editing. Mol Ther Nucleic Acids. 2017 Jun 16;7:211-222. doi: 10.1016/j.omtn.2017.04.001. Epub 2017 Apr 8. PMID: 28624197; PMCID: PMC5415201.

[2] Ma H, Marti-Gutierrez N, et al. Correction of a pathogenic gene mutation in human embryos. Nature. 2017 Aug 24;548(7668):413-419. doi: 10.1038/nature23305. Epub 2017 Aug 2. PMID: 28783728.

[3] Perez-Pinera P, Kocak DD, Vockley CM, Aet al. RNA-guided gene activation by CRISPR-Cas9-based transcription factors. Nat Methods. 2013 Oct;10(10):973-6. doi: 10.1038/nmeth.2600. Epub 2013 Jul 25. PMID: 23892895; PMCID: PMC3911785.