ArticlePlant biotechnology journal2020
Generation of a multiplex mutagenesis population via pooled CRISPR-Cas9 in soya bean.
Article in Plant biotechnology journal, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 106 papers.
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Who cites it
106 citing papers in PubMed.
- Calcium-dependent protein kinase signalling in plant immune responses: current status and future prospects.Planta · 2026Review
- Targeted allelic diversification of flowering genes via CRISPR-Cas enables the development of multiple early-flowering soybean lines.TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik · 2026Article
- Generation of Herbicide-Resistant Alfalfa Germplasm via Prime Editing.Plant biotechnology journal · 2026Article
- AI-Guided DNA-Free and Genotype-Independent Genome Editing for Soybean Improvement.Plants (Basel, Switzerland) · 2026Review
- Single-Cell CRISPR: An Efficient Strategy for Decoding Plant Cis-Regulatory Complexity.Plant biotechnology journal · 2026Review
- A small-scale CRISPR mutant library in rapeseed of commercial cultivar Zhongshuang 11.Horticulture research · 2026Article
- Efficient prime editors for heritable multiplex precision genome editing in soybean.Nature plants · 2026Article
- GmRPS5 Promoter-Driven CRISPR/LbCas12a Efficiently Generates Soybean Sextuple Mutants.Plant biotechnology journal · 2026Article
- Glycine max Jumonji C domain-containing proteins JMJ19/20 exhibit endopeptidase activity and interact with LUXs to mediate flowering time.Nucleic acids research · 2026Article
- Multiplex gene editing drives revolution in crop breeding: overlaid editing of multiple genes and customization of complex traits.Advanced biotechnology · 2026Review
- Soybean Auxin Transporter PIN3 Regulates Nitrate Acquisition to Improve Nitrogen Use and Seed Traits.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Editing a gibberellin receptor gene improves yield and nitrogen fixation in soybean.Journal of integrative plant biology · 2026Article
- CRISPR/Cas9-mediated mutagenesis ofFrontiers in plant science · 2026Article
- Advances in gene editing for legume improvement: technologies, progress, and prospects.Frontiers in genome editing · 2026Review
- CRISPR/Cas9 in soybean research: a measured leap toward genetic refinement.Frontiers in genome editing · 2026Review
- A flanking-nicks prime editor (FLICK-PE) system to boost prime editing in dicots.Nature communications · 2025Article
- A 14-3-3 modulator of seed weight and quality for unlocking the yield potential of soybean.Nature communications · 2025Article
- A CRISPR-Cas9 library to target putative redundant gene sets facilitates their functional exploration in grain development in rice.BMC plant biology · 2025Article
- CRISPR-Based genome editing in pulses: current approaches, challenges, and future prospects.Plant molecular biology · 2025Review
- Turbo-charging crop improvement: harnessing multiplex editing for polygenic trait engineering and beyond.The Plant journal : for cell and molecular biology · 2025Review
46 more citing papers are in PubMed but not listed here.
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Authors and funding
13 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The output of genetic mutant screenings in soya bean [Glycine max (L.) Merr.] has been limited by its paleopolypoid genome. CRISPR-Cas9 can generate multiplex mutants in crops with complex genomes. Nevertheless, the transformation efficiency of soya bean remains low and, hence, remains the major obstacle in the application of CRISPR-Cas9 as a mutant screening tool. Here, we report a pooled CRISPR-Cas9 platform to generate soya bean multiplex mutagenesis populations. We optimized the key steps in the screening protocol, including vector construction, sgRNA assessment, pooled transformation, sgRNA identification and gene editing verification. We constructed 70 CRISPR-Cas9 vectors to target 102 candidate genes and their paralogs which were subjected to pooled transformation in 16 batches. A population consisting of 407 T0 lines was obtained containing all sgRNAs at an average mutagenesis frequency of 59.2%, including 35.6% lines carrying multiplex mutations. The mutation frequency in the T1 progeny could be increased further despite obtaining a transgenic chimera. In this population, we characterized gmric1/gmric2 double mutants with increased nodule numbers and gmrdn1-1/1-2/1-3 triple mutant lines with decreased nodulation. Our study provides an advanced strategy for the generation of a targeted multiplex mutant population to overcome the gene redundancy problem in soya bean as well as in other major crops.
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