파지 기반 연속/비연속 단백질 유도진화 기술을 이용한 크리스퍼 유전자 편집기술의 고도화

초록

Directed evolution has become an essential approach for engineering proteins with tailored properties, but traditional round-based workflows are slow and labor-intensive. Phageassisted continuous evolution (PACE) and phage-assisted noncontinuous evolution (PANCE) overcome these limitations by coupling mutation, selection, and replication within bacteriophage- driven evolutionary circuits. These platforms enable rapid accumulation of beneficial variants and allow precise control of selection pressure through chemostat–lagoon or serial passaging systems. Recent applications of PACE/PANCE have accelerated the optimization of CRISPR genome editing technologies. Engineered Cas9 variants with expanded PAM ranges, next-generation adenine and cytosine base editors, and compact prime editors have been developed using these systems. Furthermore, PACE/PANCE has been extended to complex editing modalities, including CRISPR-associated transposases (CAST) for double-strand break-free integration and recombinaseassisted large-DNA insertion strategies such as PASTE and PASSIGE. This review summarizes the principles of PACE/ PANCE and highlights their impact on advancing CRISPRbased genome editing tools, along with perspectives on future integration with AI-driven protein design.

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