Strategies for High-Efficiency Mutation Using the CRISPR/Cas System
February 2022
in “
Frontiers in Cell and Developmental Biology
”
TLDR Improving CRISPR/Cas systems can make gene editing more efficient and precise.
The document reviewed strategies to enhance the efficiency and specificity of the CRISPR/Cas system, focusing on optimizing single-guide RNA design, using Cas protein variants, and determining optimal ribonucleoprotein concentrations. It discussed the advantages of CRISPR over traditional gene-editing tools and emphasized computational tools for sgRNA design and DNA-RNA chimera to reduce off-target effects. The review highlighted the importance of balancing nonhomologous end joining and homology-directed repair pathways, synchronizing Cas9 activity with the cell cycle, and using anti-CRISPR proteins to improve accuracy. Techniques like the S1mplex strategy and HUH-tag were noted for boosting HDR efficiency, and various delivery systems were evaluated for their efficiency and safety. The document also explored the potential of cell-penetrating peptides and abiotic delivery systems to improve cellular uptake. Despite significant progress, challenges like off-target effects and delivery inefficiencies remained, with ongoing development needed to optimize CRISPR technology for practical applications.
