ArticlePlant cell reports2017
Efficient targeted multiallelic mutagenesis in tetraploid potato (Solanum tuberosum) by transient CRISPR-Cas9 expression in protoplasts.
Article in Plant cell reports, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 185 papers, 1 of them a synthesis that pooled it.
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Who cites it
185 citing papers in PubMed, 1 synthesis or guideline pooled it.
- Genome editing for vegetable crop improvement: Challenges and future prospects.Frontiers in genetics · 2022Pooled it
- CRISPR RNP-Mediated Transgene-Free Genome Editing in Plants: Advances, Challenges and Future Directions for Tree Species.Plant, cell & environment · 2026Review
- Genome Editing in Solanaceae: Harnessing CRISPR-Cas Technology for Precision Crop Improvement.Plants (Basel, Switzerland) · 2026Review
- Genome Editing in Root and Tuber Crop Development in Sub-Saharan Africa.Plant-environment interactions (Hoboken, N.J.) · 2026Review
- Targeting a conserved functional motif in the PDS gene enables efficient CRISPR/Cas9 editing in banana.Scientific reports · 2026Article
- Protoplast-Based Functional Genomics and Genome Editing: Progress, Challenges and Applications.Plant, cell & environment · 2026Review
- Recent advances in generation of doubled haploid plants for genetic improvement in solanaceous vegetable crops.Planta · 2026Review
- Glycoalkaloid-Free Starch Potatoes Generated by CRISPR/Cas9-Mediated Mutations of Genes in the Glycoalkaloid Biosynthesis Pathway Enable More Sustainable Uses of By-Products From Starch Production.Plant biotechnology journal · 2026Article
- From Lab to Field: CRISPRing Major CultivatedInternational journal of molecular sciences · 2026Review
- A Stable and Rapid Protoplast-Based CRISPR/Cas9-Ribonucleoprotein Complex Screening System for Octoploid Strawberry (Plants (Basel, Switzerland) · 2026Article
- CRISPR-Cas technologies for precision genome editing in plants: advances, applications, and future perspectives.Frontiers in plant science · 2026Review
- Impedance flow cytometry for rapid quality assessment of protoplast cultures.Plant methods · 2025Article
- A novel recombinant CRISPR/Cas9 vector system for genome editing in plants.Transgenic research · 2025Article
- CRISPR-Cas Gene Editing Technology in Potato.International journal of molecular sciences · 2025Review
- Integrating genome editing with omics, artificial intelligence, and advanced farming technologies to increase crop productivity.Plant communications · 2025Review
- CRISPR mediated gene editing for economically important traits in horticultural crops: progress and prospects.Transgenic research · 2025Review
- Advancing vegetable genetics with gene editing: a pathway to food security and nutritional resilience in climate-shifted environments.Functional & integrative genomics · 2025Review
- The giant genome of lily provides insights into the hybridization of cultivated lilies.Nature communications · 2025Article
- Enhancing quality and climate resilient traits in vegetatively propagated polyploids: transgenic and genome editing advancements, challenges and future directions.Frontiers in genetics · 2025Review
- CRISPR/Cas9 editing ofFrontiers in plant science · 2025Article
125 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
key messageAltered starch quality with full knockout of GBSS gene function in potato was achieved using CRISPR-Cas9 technology, through transient transfection and regeneration from isolated protoplasts. Site-directed mutagenesis (SDM) has shown great progress in introducing precisely targeted mutations. Engineered CRISPR-Cas9 has received increased focus compared to other SDM techniques, since the method is easily adapted to different targets. Here, we demonstrate that transient application of CRISPR-Cas9-mediated genome editing in protoplasts of tetraploid potato (Solanum tuberosum) yielded mutations in all four alleles in a single transfection, in up to 2 % of regenerated lines. Three different regions of the gene encoding granule-bound starch synthase (GBSS) were targeted under different experimental setups, resulting in mutations in at least one allele in 2-12 % of regenerated shoots, with multiple alleles mutated in up to 67 % of confirmed mutated lines. Most mutations resulted in small indels of 1-10 bp, but also vector DNA inserts of 34-236 bp were found in 10 % of analysed lines. No mutations were found in an allele diverging one bp from a used guide sequence, verifying similar results found in other plants that high homology between guide sequence and target region near the protospacer adjacent motif (PAM) site is essential. To meet the challenge of screening large numbers of lines, a PCR-based high-resolution fragment analysis method (HRFA) was used, enabling identification of multiple mutated alleles with a resolution limit of 1 bp. Full knockout of GBSS enzyme activity was confirmed in four-allele mutated lines by phenotypic studies of starch. One remaining wild-type (WT) allele was shown sufficient to maintain enough GBSS enzyme activity to produce significant amounts of amylose.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.