ArticlePlanta2019
CRISPR/Cas9-mediated knockout of the RDR6 gene in Nicotiana benthamiana for efficient transient expression of recombinant proteins.
Article in Planta, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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Who cites it
26 citing papers in PubMed, 44 citations in OpenAlex.
- Bioluminescent sentinel plants enable autonomous diagnostics of viral infections.Nature communications · 2026Article
- Editing of an antiviral host factor boosts plant growth and yield of plant viral vector-mediated heterologous protein expression.Plant biotechnology journal · 2026Article
- Recovery of RNA-dependent RNA polymerase 6 gene-knockout phenotypes in Nicotiana benthamiana via in vivo generation of inverted repeat construct of the trans-acting short interference RNA3 sequence.The Plant journal : for cell and molecular biology · 2025Article
- Developing a multi-modular assembled prime editing (mPE) system improved precise multi-base insertion efficiency in dicots.Journal of advanced research · 2025Article
- Plant-made pharmaceuticals.Plant biotechnology (Tokyo, Japan) · 2024Article
- Optimizing Promoters and Subcellular Localization for Constitutive Transgene Expression in Marchantia polymorpha.Plant & cell physiology · 2024Article
- Improving transient protein expression in agroinfiltrated Nicotiana benthamiana.The New phytologist · 2024Review
- A novel ilarvirus protein CP-RT is expressed via stop codon readthrough and suppresses RDR6-dependent RNA silencing.PLoS pathogens · 2024Article
- Improving transient expression inFrontiers in plant science · 2024Article
- Crop genome editing through tissue-culture-independent transformation methods.Frontiers in genome editing · 2024Review
- CRISPR/Cas9-gene editing approaches in plant breeding.GM crops & food · 2023Review
- Green Biologics: Harnessing the Power of Plants to Produce Pharmaceuticals.International journal of molecular sciences · 2023Review
- Article
- Seed transmission of raspberry bushy dwarf virus is blocked in Nicotiana benthamiana plants by preventing virus entry into the embryo from the infected embryo sac and endosperm.Archives of virology · 2023Article
- Differential Effects of RNA-Dependent RNA Polymerase 6 (RDR6) Silencing on New and Old World Begomoviruses inViruses · 2023Article
- Functional Characterization ofPlants (Basel, Switzerland) · 2023Article
- Increased rates of gene-editing events using a simplified RNAi configuration designed to reduce gene silencing.Plant cell reports · 2022Article
- CRISPR/Cas9-mediated knockout of the DCL2 and DCL4 genes in Nicotiana benthamiana and its productivity of recombinant proteins.Plant cell reports · 2022Article
- Improving Protein Quantity and Quality-The Next Level of Plant Molecular Farming.International journal of molecular sciences · 2022Review
- CRISPR/Cas9-Mediated Genome Editing System in Insect Genomics and Pest Management.Methods in molecular biology (Clifton, N.J.) · 2022Article
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
MAIN
conclusionRDR6 gene knockout Nicotiana benthamiana plant was successfully produced using CRISPR/Cas9 technology. The production of recombinant proteins in plants has many advantages, such as safety and reduced costs. However, there are several problems with this technology, especially low levels of protein production. The dysfunction of the RNA silencing mechanism in plant cells would be effective to improve recombinant protein production because the RNA silencing mechanism efficiently degrades transgene-derived mRNAs. Therefore, to overcome this problem, clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 technology was used to develop RNA silencing-related gene knockout transgenic Nicotiana benthamiana. We successfully produced RNA-dependent RNA polymerase 6 (RDR6), one of the most important components of the RNA silencing mechanism-knockout N. benthamiana (ΔRDR6 plants). The ΔRDR6 plants had abnormal flowers and were sterile, as with the Arabidopsis RDR6 mutants. However, a transient gene expression assay showed that the ΔRDR6 plants accumulated larger amounts of green fluorescent protein (GFP) and GFP mRNA than the wild-type (WT) plants. Small RNA sequencing analysis revealed that levels of small interfering RNA against the GFP gene were greatly reduced in the ΔRDR6 plants, as compared to that of the WT plants. These findings demonstrate that the ΔRDR6 plants can express larger amounts of recombinant proteins than WT plants and, therefore, would be useful for recombinant protein production and understanding the contributions of RDR6 to genetic and physiological events in plants.
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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.