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pp.231-235
Despite their historical use, studies on the genetic functions of mushrooms and varietal improvement via biomolecular techniques are limited compared to other organisms. Recent advancements in CRISPR/Cas9 have enabled precise genetic modifications in mushrooms, with RNP-based systems offering high editing efficiency without foreign gene insertion. In this study, we optimized gene-editing conditions for Ganoderma lucidum (Yongji 2) by utilizing RNP/nanoparticle complexes to enhance efficiency. The optimal conditions included a 0.2 M sorbitol buffer (pH 7.0) and a protoplast-to-complex ratio of 10:1. Among eight gRNAs designed for the catA gene, three were identified with high activity, and PEG-mediated transformation resulted in successful gene edits, primarily involving 1 bp deletions. The editing efficiency reached 7–8%, demonstrating that nanoparticle-supported RNP systems are effective for marker-free gene editing in mushrooms. These findings highlight a promising approach for advancing genetic research and varietal improvement in G. lucidum and other mushroom species.
서 론
재료 및 방법
균주 및 배양
원형질체 분리
gRNA의 디자인과 RNP 생산
RNP/나노파티클 복합체 형성 최적화
원형질체, RNP/나노파티클 혼합비 최적화
결과 및 고찰
영지 RNP/나노파티클 복합체 유전자 교정 최적화
RNP/나노파티클 복합체를 활용한 영지 유전자 교정
적 요
감사의 글
REFERENCES
