ArticleBMC plant biology2020
A stable DNA-free screening system for CRISPR/RNPs-mediated gene editing in hot and sweet cultivars of Capsicum annuum.
Article in BMC plant biology, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 36 papers.
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Who cites it
36 citing papers in PubMed, 87 citations in OpenAlex.
- 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
- From Lab to Field: CRISPRing Major CultivatedInternational journal of molecular sciences · 2026Review
- Tiny but powerful: protoplast isolation inFrontiers in plant science · 2026Article
- Highly efficient transgene-free ErCas12a RNP-protoplast genome editing and single-cell regeneration in Nicotiana benthamiana for glyco-engineering.Plant biotechnology journal · 2026Article
- Genetic transformation and genome editing of flax (Frontiers in plant science · 2026Review
- The Enhancement of Fungal Disease Resistance in Major Staple Crops Using CRISPR-Cas Technology.Genes · 2025Review
- Genome Editing by Grafting.International journal of molecular sciences · 2025Review
- A visual monitoring DNA-free multi-gene editing system excised via LoxP::FRT/FLP in poplar.Plant biotechnology journal · 2025Article
- Strategies to develop climate-resilient chili peppers: transcription factor optimization through genome editing.Planta · 2025Review
- Development of a CRISPR/Cas9 RNP-mediated genetic engineering system in Paecilomyces variotii.Journal of microbiology (Seoul, Korea) · 2025Article
- The mitochondrial genome data of theData in brief · 2025Article
- Molecular and genomic insights into viral resistance inFrontiers in plant science · 2025Review
- Highly efficient CRISPR/Cas9-RNP mediatedPlant signaling & behavior · 2024Article
- Enhancing genetic modification in recalcitrant plants: An investigation in chili (Plant biotechnology (Tokyo, Japan) · 2024Article
- Article
- CRISPR/Cas9 based genome editing of Phytoene desaturase (PDS) gene in chilli pepper (Capsicum annuum L.).Journal, genetic engineering & biotechnology · 2024Article
- Review
- Elucidating the callus-to-shoot-forming mechanism in Capsicum annuum 'Dempsey' through comparative transcriptome analyses.BMC plant biology · 2024Article
- Harnessing CRISPR/Cas9 for Enhanced Disease Resistance in Hot Peppers: A Comparative Study onInternational journal of molecular sciences · 2023Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundDNA-free, clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein (Cas) ribonucleoprotein (RNP)-based genome editing is a simple, convincing, and promising tool for precision crop breeding. The efficacy of designed CRISPR-based genome editing tools is a critical prerequisite for successful precision gene editing in crops.
resultsThis study demonstrates that soil-grown leaf- or callus-derived pepper protoplasts are a useful system for screening of efficient guide RNAs for CRISPR/Cas9 or CRISPR/Cas12a (Cpf1). CRISPR/Cas9 or Cpf1 were delivered as CRISPR/RNP complexes of purified endonucleases mixed with the designed single guide RNA, which can edit the target gene, CaMLO2 in two pepper cultivars with whole genome sequenced, Capsicum annuum 'CM334' and C. annuum 'Dempsey'. The designed guide RNAs (sgRNAs for Cas9 or crRNAs for Cpf1) are conserved for CaMLO2 in both CM334 and Dempsey and cleave CaMLO2 in vitro. CRISPR/Cas9- or /Cpf1-RNP complexes were transfected into purely isolated protoplasts of the hot pepper CM334 and sweet pepper Dempsey by PEG-mediated delivery. Targeted deep sequencing analysis indicated that the targeted CaMLO2 gene was differentially edited in both cultivars, depending on the applied CRISPR/RNPs.
conclusionsPepper protoplast-based CRISPR guide-RNA selection is a robust method to check the efficacy of designed CRISPR tools and is a prerequisite for regenerating edited plants, which is a critical time-limiting procedure. The rapid and convincing selection of guide RNA against a target genome reduces the laborious efforts for tissue culture and facilitates effective gene editing for pepper improvement.
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