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Objective <p>Plant transformation frequently results in large quantities of regenerant plant materials which must undergo screening prior to advancement into subsequent experiments. In-depth sequencing of such quantities for detection of editing events is costly, and preliminary selection of mutant lines based on phenotypic impacts can cause significant delays. This study aimed to develop a rapid, high-throughput workflow for early detection and genotyping of CRISPR/Cas9-mediated edits in strawberry.</p> Results <p>High-resolution analysis reliably identified lines containing distinct mutation profiles, and subsequent Amplicon sequencing genotyping confirmed the presence of editing events at the targeted sites, with editing efficiencies varying among subgenomes of octoploid strawberry. This workflow offers a scalable, low-cost solution for early detection and prioritization of genome-edited cultivated strawberry lines.</p>

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Rapid screening of genome edited strawberry (Fragaria ×ananassa) regenerants using high-resolution melting analysis followed by Amplicon sequencing

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年4月24日

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  • Kaitlyn Vondracek,
  • Man Bo Lee,
  • Tie Liu,
  • Seonghee Lee

摘要

Objective

Plant transformation frequently results in large quantities of regenerant plant materials which must undergo screening prior to advancement into subsequent experiments. In-depth sequencing of such quantities for detection of editing events is costly, and preliminary selection of mutant lines based on phenotypic impacts can cause significant delays. This study aimed to develop a rapid, high-throughput workflow for early detection and genotyping of CRISPR/Cas9-mediated edits in strawberry.

Results

High-resolution analysis reliably identified lines containing distinct mutation profiles, and subsequent Amplicon sequencing genotyping confirmed the presence of editing events at the targeted sites, with editing efficiencies varying among subgenomes of octoploid strawberry. This workflow offers a scalable, low-cost solution for early detection and prioritization of genome-edited cultivated strawberry lines.