Post 2000, a number of advances in recombinant DNA technology has enabled scientists to develop the means to use CRISPR sequences to edit mammalian genomes. This resulted in the establishment of the CRISPR/Cas9 genome editing system. This gene editing tool kit comprises of a short guide RNA (gRNA) sequence, a DNA specific CRISPR RNA (crRNA) sequence and the Cas9 protein. The gRNA sequence is a partially conserved palindromic sequence, which directs the Cas9 protein to a specific location in the genome. This step brings about a double strand break in the genomic DNA, disrupting the targeted gene segment. The DNA-specific crRNA is then used to edit the target sequence; this may involve a deletion, an insertion or a replacement of oligonucleotide sequences. The nuclease protein Cas9 facilitates the gene replacement phenomenon via homologous recombination. In special cases, components such as trans-activating CRISPR RNA (trRNA) are also added to the mix to assist the gene editing process.
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