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CRISPR-based technologies for large DNA insertions
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DOI：10.1016/j.tibtech.2026.04.004.png)
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Abstract
En 中文
Large DNA insertion technologies are critical for advancing medicine, biotechnology, and agriculture. CRISPR-Cas9 technology is limited by double-stranded breaks and cargo-size constraints. New hybrid systems combine CRISPR programmability with DNA integration technologies. Twin prime editing with Bxb1, programmable addition via site-specific targeting element, prime-editing-assisted site-specific integrase gene editing, and prime-editing-mediated recombination of opportune target achieve targeted, large, and double-stranded break-free insertions. Evolved recombinases (evoBxb1 and eeBxb1) dramatically improve efficiency and specificity. Continued innovation in protein engineering, directed evolution, and artificial intelligence design will drive next-generation genome-editing technologies.
Keywords:
CRISPR-Cas
large DNA insertion
mobile genetic elements
site-specific recombinase
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