Development of an efficient construction method for ZFNs

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With the advent of the CRISPR-Cas system, genome editing technology is now readily available to everyone. Compared to CRISPR-Cas, TALEN (Transcription activator-like effector nuclease) is a bit more complicated to manufacture than CRISPR-Cas, but it has a good track record and is worth using.However, commercial use of TALEN requires payment of a license fee, which is also an obstacle to its use. However, Zinc-Finger Nuclease (ZFN), a “first generation” genome editing tool, has already reached the maturity of its main patents (Scott, Nat Biotechnol, 2005), and is again attracting attention as a tool for commercial use. However, compared to CRISPR-Cas and TALEN, ZFNs have not been widely used due to a number of challenges such as difficulty in designing arbitrary nucleotide sequence targets, months of tedious construction work, and low success rate of construction.

We have experience in establishing ZFN construction methods (Ochiai et al., Genes Cells, 2010; Ochiai et al., PNAS, 2012) and aim to develop more efficient ZFN construction methods by weaving the latest findings. This is very important not only for commercial applications but also for academic purposes to better understand the DNA recognition mechanism of zinc-finger proteins, which account for more than half of the transcription factors in eukaryotic genomes.