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GENOME EDITING - Coggle Diagram
GENOME EDITING
Programmable nucleases
Produce site-specific DNA double-strand breaks (DSBs), which enhance the efficiency of homologous recombination and/or trigger error-prone non-homologous end-joining (NHEJ), which leads to targeted mutagenesis
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Gene disruption: through error prone NHEJ, which gives rise to small insertions and deletions (indels) at the nuclease target site. Indels cause frameshifts in the coding region, which disrupt genetic information and result in gene knockout.
Gene insertion: plasmid DNA or integrating viral vectors are used. Engineered nucleases enhance the efficiency of homologous recombination in a site-specific manner. The nuclease is co-transfered with a targeting vector, in which the genetic segment to be incorporated is flanked by homology arms with sequences that are identical to those near the target region. Alternatively, it defined overhangs are generated by nucleases, then specific sequences can be inserted into the target region by NHEJ-mediated ligation.
Gene correction and point mutagenesis: through co-delivery or programmable nucleases and targeting vectors or single-strand oligodeoxynucleotides (ssODNs).
Chromosomal rearrangements: the repair of two concurrent DSBs induced by programmable nucleases can give rise to chromosomal rearrangements or structural variations in a targeted manner. Deletions, duplications and inversions of up to a few Mbp of chromosomal segments have been achieved using ZFNs, TALENs or RGENs.
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