ArticleNature protocols2020
Manipulating gene translation in plants by CRISPR-Cas9-mediated genome editing of upstream open reading frames.
Article in Nature protocols, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 47 papers.
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Who cites it
47 citing papers in PubMed, 88 citations in OpenAlex.
- Coordinating structure-guided design and cis-regulatory optimization to engineer elite herbicide-tolerance alleles in crops.Nature plants · 2026Article
- CRISPR-Cas9-mediated uATG introduction in the 5'UTR of the Uox gene for hyperuricemia mouse models: implications for gout and metabolic disorders.Science China. Life sciences · 2026Article
- A Fluorescence-Based Transient Expression Assay for the Analysis of Upstream Open Reading Frames in Plants.Plant direct · 2026Article
- Maize Anther Development Involves Translated Open Reading Frames From 3' Untranslated Regions.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Targeted BDNF upregulation via upstream open reading frame disruption.Molecular therapy : the journal of the American Society of Gene Therapy · 2026Article
- Not just Rubisco: integrated strategies to overcome the biochemical limitations of CFrontiers in plant science · 2026Review
- CRISPR/Cas9-mediated uORF engineering enhances tanshinone biosynthesis inHorticulture research · 2026Article
- Engineering non-coding DNA Elements in Rice: an Elegant Approach To fine-tune Agronomical Advantageous Traits.Rice (New York, N.Y.) · 2025Review
- The Potential of NGTs to Overcome Constraints in Plant Breeding and Their Regulatory Implications.International journal of molecular sciences · 2025Review
- Variations in the OsGGP uORF Fine-Tune Vitamin C Content and Confer Resistance to Osmotic Stress in Rice.Rice (New York, N.Y.) · 2025Article
- Antioxidant Defense Systems in Plants: Mechanisms, Regulation, and Biotechnological Strategies for Enhanced Oxidative Stress Tolerance.Life (Basel, Switzerland) · 2025Review
- Recent advances in improving yield and immunity through transcription factor engineering.Journal of integrative plant biology · 2025Review
- Through the lens of bioenergy crops: advances, bottlenecks, and promises of plant engineering.The Plant journal : for cell and molecular biology · 2025Review
- Article
- Gene Editing inJournal of fungi (Basel, Switzerland) · 2025Review
- Plant engineering: advances, bottlenecks, and promise.The Plant journal : for cell and molecular biology · 2025Article
- Genetic engineering, including genome editing, for enhancing broad-spectrum disease resistance in crops.Plant communications · 2025Review
- Increasing Rubisco as a simple means to enhance photosynthesis and productivity now without lowering nitrogen use efficiency.The New phytologist · 2025Review
- Combining AI and new genomic techniques to 'fine-tune' plants: challenges in risk assessment.Frontiers in plant science · 2025Article
- CRISPR activation: identifying and using novel genes for plant disease resistance breeding.Frontiers in genome editing · 2025Review
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Gene expression is regulated by multiple processes, and the translation of mRNAs into proteins is an especially critical step. Upstream open reading frames (uORFs) are widespread cis-elements in eukaryotic genes that usually suppress the translation of downstream primary ORFs (pORFs). Here, we describe a protocol for fine-tuning gene translation in plants by editing endogenous uORFs with the CRISPR-Cas9 system. The method we present readily yields transgene-free uorf mutant offspring. We provide detailed protocols for predicting uORFs and testing their effects on downstream pORFs using a dual-luciferase reporter system, designing and constructing single guide RNA (sgRNA)-Cas9 vectors, identifying transgene-free uorf mutants, and finally comparing the mRNA, protein and phenotypic levels of target genes in uorf mutants and controls. Predicting uORFs and confirming their effects in protoplasts takes only 2-3 weeks, and transgene-free mutants with edited target uORFs controlling different levels of pORF translation can be obtained within 4 months. Unlike previous methods, our strategy achieves fine-tuning of gene translation in transgene-free derivatives, which accelerates the analysis of gene function and the improvement of crop traits.
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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.