Advantages
The primary advantage of this technique is its ability to eliminate any foreign DNA from the genome after the mutagenesis process. This ensures there are no selectable markers or exogenous sequences used for targeting left in the genome that may cause unforeseen effects.
The delitto perfetto technique is also simpler compared to other methods for in vivo site-directed mutagenesis. Other methods require multiple cloning steps and extensive DNA sequencing to confirm mutagenesis, which is often a complicated and inefficient process.
There is great flexibility in this approach because after the CORE cassette is inserted (see Method Overview for details), multiple mutations in the gene of interest can be made easily and quickly.
This method can be applied to other organisms where homologous recombination is efficient, such as the moss Physcomitrella patens, DT40 chicken cells, or E. coli. In addition, human genes can be studied and similarly genetically manipulated in yeast by using yeast artificial chromosomes (YACs).
Read more about this topic: Delitto Perfetto
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