In every illustration and available resources it says that the two major pathways for repair of DSB is either homologous recombination (HR) or nonhomologous end joining (NHEJ) in crispr cas9 system
how does this possible in artificially introduced systems (e.g in plants)? Do they inherit required enzymes / proteins naturally for this kind of situation?
As I've understood if I introduced gRNA to plant for crispr cas9 system, it will create the DSB, if however this break repair by HR, again DSB will create by the expressing gRNA crispr cas9 system....Unless if there will be a insertion / deletion / mutation this process will repeat like a cycle??
Also, suppose that we introduce Homologous repair template along with the Cas9-sgRNA , So is this mechanism that much effective and efficiency of screening requires condition of our experiment in plants.
I wonder what actually happen actually? Can some one explain this :)
1. When CRISPR-cas9 is introduced into plant cells, it targets and causes 'DSB' on a specific endogenous plant gene/allele (a gene/allele you have chosen). After the DSB is created, host cellular repairing system (which provides necessary enzymes) will kick in to repair the DSB.
2. After DSB is created and repaired by NHEJ, the DNA sequence of the site can change (some add a few bases, some delete bases). The change of the sequences cannot be re-recognized by sgRNA/Cas9 nuclease as a target.
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