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<p>Plant genome editing is moving from recipe-driven optimization to evidence-based engineering. Here we synthesize DNA-free CRISPR workflows in Solanaceae (tomato as the primary model, with cross-references to pepper and potato) to define quantitative, reproducible benchmarks from transformation to multi-environment field testing and regulatory readiness. Using a structured literature screening and coding scheme, we map (i) delivery strategies (RNPs, transient expression, ribonucleoprotein–protoplast/meristem approaches), (ii) validation tiers integrating on-target integrity (amplicon- and long-read WGS), off-target discovery (CHANGE-seq/capture-seq), and line segregation, and (iii) decision rules for advancement, including power-informed non-inferiority margins for agronomic performance in multi-environment trials. We distill a three-table toolkit: Editing mode × evidence matrix; MET design with sample-size guidance for genotype × environment; and a jurisdiction-agnostic compliance checklist focusing on DNA-free status, molecular characterization, and phenotypic equivalence. Case vignettes in tomato illustrate how benchmark thresholds (e.g., minimum read-depth/coverage for structural variant screening; concordance across orthogonal assays) de-risk scale-up while preserving generalizability. The review concludes with actionable templates (protocol fields, reporting items, and acceptance gates) to convert lab-level edits into field-ready, auditable outcomes.</p>

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DNA-free genome editing in Solanaceae: an evidence-based roadmap from transformation to field and regulatory readiness

  • 期刊论文

  • 发表日期：

    2026年2月24日

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  • Ying Qin,
  • Wenwen Si,
  • Chengchuang Huang

摘要

Plant genome editing is moving from recipe-driven optimization to evidence-based engineering. Here we synthesize DNA-free CRISPR workflows in Solanaceae (tomato as the primary model, with cross-references to pepper and potato) to define quantitative, reproducible benchmarks from transformation to multi-environment field testing and regulatory readiness. Using a structured literature screening and coding scheme, we map (i) delivery strategies (RNPs, transient expression, ribonucleoprotein–protoplast/meristem approaches), (ii) validation tiers integrating on-target integrity (amplicon- and long-read WGS), off-target discovery (CHANGE-seq/capture-seq), and line segregation, and (iii) decision rules for advancement, including power-informed non-inferiority margins for agronomic performance in multi-environment trials. We distill a three-table toolkit: Editing mode × evidence matrix; MET design with sample-size guidance for genotype × environment; and a jurisdiction-agnostic compliance checklist focusing on DNA-free status, molecular characterization, and phenotypic equivalence. Case vignettes in tomato illustrate how benchmark thresholds (e.g., minimum read-depth/coverage for structural variant screening; concordance across orthogonal assays) de-risk scale-up while preserving generalizability. The review concludes with actionable templates (protocol fields, reporting items, and acceptance gates) to convert lab-level edits into field-ready, auditable outcomes.