International Journal of Molecular Medical Science, 2024, Vol.14, No.4, 252-263 http://medscipublisher.com/index.php/ijmms 255 fully understood, and there are concerns about the heritability of genetic modifications and their impact on future generations (Barman et al., 2020). These challenges underscore the need for thorough preclinical testing and stringent regulatory oversight to ensure the safety and efficacy of CRISPR/Cas9-based therapies for AD. Figure 2 Strategy and manipulation of the amyloid pathway by CRISPR/Cas9 editing (Adopted from Sun et al., 2018) Image caption: The study utilized CRISPR/Cas9 technology to target the C-terminal of the amyloid precursor protein (APP), which is associated with Alzheimer's disease, and successfully achieved gene editing; a: This panel shows the sgRNA target sequence and its location within the genome, with antibody Y188 used to confirm successful editing of the APP C-terminal; b: This panel illustrates that the fluorescence signal of APP in neuroblastoma cells treated with CRISPR/Cas9 is significantly reduced, indicating successful editing of APP; c and d: These panels further confirm, using immunoblotting, the reduction of the APP C-terminal fragment (APP-CTF) following CRISPR treatment, while the N-terminal signal remains unchanged, indicating that the editing specifically affects the APP C-terminal; e: This panel shows the time course of the editing effect, with a gradual decrease in APP-CTF levels after editing; f: This panel demonstrates the efficient editing of mouse APP at the expected target site, validated by deep genome sequencing; g: This panel compares the APP target sequences in humans and mice, noting minor differences but showing that effective editing can still be achieved using human-specific sgRNA; h-i: These panels show that APP C-terminal editing was also successfully achieved in human iPSC-derived neurons, promoting an increase in neuroprotective α-cleavage while reducing β-cleavage products, confirming the effectiveness of the CRISPR/Cas9 strategy at the cellular level (Adapted from Sun et al., 2018) Gene therapy targeting the amyloid-beta and tau pathways, enhancing neuroprotective proteins, and utilizing CRISPR/Cas9 technology holds great promise for the treatment of Alzheimer's disease. As research progresses,
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