CRISPR-Cas9 for the genome engineering of cyanobacteria and succinate production (Li et al, 2016).
CRISPR-Cas9
Figure 5. CRISPR-Cas system. (PNA Bio, 2014)
CRISPR-Cas9 is a genome editing tool. It is faster, cheaper and more accurate than previous techniques of editing DNA and has a wide range of potential applications, CRISPR-Cas9 is a unique technology that enables geneticists and medical researchers to edit parts of the genome by removing, adding or altering sections of the DNA sequence (Yourgenome, 2015).
Figure 6. CRISPR-Cas9 in action. (McGovern Institute for Brain Research at MIT, 2014)
The CRISPR-Cas9 system consists of two key molecules that introduce a change (mutation) into the DNA. These are: the enzyme called Cas9. This acts as a pair of “molecular scissors” that can cut the two strands of DNA at a specific location in the genome so that bits of DNA can then be added or removed. The second is a piece of RNA called guide RNA (gRNA). This consists of a small piece of pre-designed RNA sequence located within a longer RNA scaffold. The scaffold part binds to DNA and the pre-designed sequence “guides” Cas9 to the right part of the genome. This makes sure that the Cas9 enzyme cuts at the right point in the genome (Yourgenome, 2015).
The guide RNA is designed to find and bind to a specific sequence in the DNA. The guide RNA has RNA bases that are complementary to those of the target DNA sequence in the genome. This means that, at least in theory, the guide RNA will only bind to the target sequence and no other regions of the genome. The Cas9 follows the guide RNA to the same location in the DNA sequence and makes a cut across both strands of the DNA. At this stage the cell recognizes that the DNA is damaged and tries to repair it (Yourgenome, 2015).
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This video is about the genome editing whit CRISPR-Cas9, presents the technique in the most simply form. Is from McGovern Institute for Brain Research at MIT propriety.