Precise Genetics: New CRISPR Method Enables Efficient DNA Modification
Published:22 Sep.2024 Source:University of Basel
The research group led by Prof. Markus Affolter at the Biozentrum, University of Basel, has developed a new method that further improves the existing CRISPR/Cas technologies: it allows a more precise and seamless introduction of tags into proteins at the gene level. This technology could significantly improve research on proteins in living organisms and opens up new possibilities for medical research. With the revolutionary CRISPR/Cas technology, the DNA of living organisms can be precisely altered. Using a guide RNA that recognizes a specific DNA sequence, Cas9 protein is recruited to that sequence and cuts the DNA. This targeted cut allows the DNA to be repaired or altered at this specific location.
Prof. Markus Affolter's team at the Biozentrum, University of Basel, has now developed a new method called SEED/Harvest in the fruit fly (Drosophila melanogaster). This method combines the CRISPR-Cas9 technique with the Single-Strand Annealing (SSA) repair pathway, enabling genome-wide changes to be carried out more efficiently and without leaving unwanted scars. The SEED/Harvest method proceeds in two steps. In a first step, the researchers introduced a marker gene into the desired DNA site within a protein-coding region. This marker is placed at the targeted location and is used to isolate successful modifications. In a second step, the marker is excised and the DNA breakpoints are repaired by the Single-Strand Annealing (SSA) repair pathway. The combination of both methods makes it possible to mark any desired protein in the genome without collateral damage, allowing us to study the functions of proteins in living organisms.
The SEED/Harvest method is both. It combines the most robust screening of successful insertions and all the advantages of seamless tagging. One of the advantages of the SEED/Harvest method is that proteins can be labeled in specific tissues and cell types. This opens up new possibilities for research to investigate the dynamics of proteins systematically in living cells in real-time.