Genome editing tools

I- ZFN

    Zinc finger nuclease (ZFN) is a class of synthetic restriction enzymes that can target specific genome site and produce site-specific double strand DNA cleavages. ZFNs consist of two parts: one part is called zinc-finger protein (ZFP) that recognize and bind specific genomic sites; and another part is FokI nuclease that non-specifically cuts DNA and results in double strand DNA breakage.

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II- TALEN

    Like ZFN, TALEN (Transcription Activator-Like Element Nuclease) is a class of synthetic restriction enzymes that can target specific genome site and produce site-specific double strand DNA cleavages.  TALENs also consist of two parts: one part is called transcription activator-like element (TALE) that recognize and bind specific genomic sites; and another part is FokI nuclease that non-specifically cuts DNA and results in double strand DNA breakage. 

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III- CRISPR/Cas9

    CRISPR / Cas9 genome editing technology is a new yet widely adopted genome editing technology. Unlike ZFN and TALEN, DNA sequence recognition of CRISPR / Cas9 is mediated by RNA-DNA interaction, not protein-DNA interaction. Also, the DNA cutting enzyme is Cas9 that recognizes specific RNA-DNA binding structure, not FokI that is non-specific.  CRISPR / Cas9 are much easier to design and assemble compared to ZFN and TALEN; therefore, it has become the dominant technology for genome editing.  For CRISPR / Cas9 to work, one need to design a site-specific guild RNA that binds to targeted genome site. Once a guild RNA binds DNA site, it forms a specific structure that will recruit Cas9 nuclease to the site and cause double strand DNA cleavage.   In general, CRISPR / Cas9 have higher cutting activities compared to ZFN and TALEN and much easier to be adapted to different applications.

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