I want to use CRISPR/Cas9 in a cell line to introduce a specific mutation that will permanently remain in the cells for further study, changing one base for another. If I use a transient vector for CRISPR/Cas9 to induce a stable mutation via HDR using a DNA donor, will the mutation be permanent or not? Do I need to use only a constitutive vector for this?
Adrieli Silva de Carlis Transient vector is typically adequate and preferred to prevent extended Cas9 activity. However, its efficiency may be lower due to the dominance of error-prone repair mechanisms over template-directed editing. To enhance targeted modifications in such cases, strategies like chemical promotion of HDR or suppression of NHEJ-related proteins can be employed. A constitutive vector is generally necessary only when sustained editing is required, such as for gene regulation studies.
- Badges
- Science topic
- Similar topics
- Biological Science
- Anatomy
- Tissue