Today we have access to technologies that enable us to insert genes into hosts, control gene expression and to reduce or completely inhibit the expression of certain genes. The CRISPR/Cas9 technology started emerging in 2013 and has now become an effective and relatively simple method for genome editing. It is based on bacterial antiviral immune response. However, the most common limitation within using the CRISPR/Cas9 method is the creation of DNA double stranded breaks, which are not always our goal and the possibility of off-target binding of the CRISPR/Cas9 system and DNA sequence. Consequently, the CRISPRi/dCas9 method, which enables us to decrease the expression of genes and the CRISPRa/dCas9 method, an optimised technology for overexpression of genes, were developed. For a more efficient regulation of expression of target genes, these methods are being used together as CRISPRai. This technology allows us to simultaneously repress one and activate another gene with greater precision and specificity, without changing the DNA sequence. In this work, we will look into the advantages, specific examples and the broad spectrum of usages for CRISPRai/dCas9, with accents on metabolic or genetic engineering, in biomedicine, diagnostics, targeted drug development, cell and gene therapy and in some other related fields.
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