ArticlePlant biotechnology journal2025
PAM-relaxed and temperature-tolerant CRISPR-Mb3Cas12a single transcript unit systems for efficient singular and multiplexed genome editing in rice, maize, and tomato.
Article in Plant biotechnology journal, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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16 citing papers in PubMed.
- Establishment of an efficient and PAM-relaxed LbCas12a genome editing tool in plants.The New phytologist · 2026Article
- Advances in CRISPR multiplex gene editing to map and modify stress-responsive transcription factor networks for crop improvement.Plant cell reports · 2026Review
- Genome Editing in Solanaceae: Harnessing CRISPR-Cas Technology for Precision Crop Improvement.Plants (Basel, Switzerland) · 2026Review
- Enhanced exonuclease-Cas9 systems promote multiple nucleotide deletions with higher efficiency and broader targeting scope in plants.Journal of integrative plant biology · 2026Article
- Rapid On-Site Detection ofFoods (Basel, Switzerland) · 2026Article
- Wheat's Up with CRISPR-Cas-Current Advances, Obstacles and Perspectives.International journal of molecular sciences · 2026Review
- Tuning Rice Gene Expression via In Situ Promoter Truncations Using a Prime Editing Library.Plant biotechnology journal · 2026Article
- Advances in Genome Editing for Plant Disease Resistance Breeding.Plants (Basel, Switzerland) · 2026Review
- Multiplex gene editing drives revolution in crop breeding: overlaid editing of multiple genes and customization of complex traits.Advanced biotechnology · 2026Review
- Enhancing heritable genome editing in soybean by optimizing promoter combinations for the LbCas12a system.BMC plant biology · 2026Article
- Precision breeding in a changing climate: unlocking resilience through omics and gene editing.Functional & integrative genomics · 2026Review
- Research progress on cold tolerance key genes and mechanisms in four major northern China crops.Frontiers in plant science · 2026Review
- Beyond the Edit: Integrating CRISPR-Cas Genome Editing Into the Cereal Breeding Pipeline.International journal of genomics · 2026Review
- Programmable genome engineering and gene modifications for plant biodesign.Plant communications · 2025Review
- Toward a monocot SynBio toolkit: assessing regulatory element performance and eudicot compatibility.Plant cell reports · 2025Article
- CRISPR-Cas12i confers efficient genome editing and gene regulation in plants.Plant physiology · 2025Article
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14 authors.
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Abstract
Class 2 Type V-A CRISPR-Cas (Cas12a) nucleases are powerful genome editing tools, particularly effective in A/T-rich genomic regions, complementing the widely used CRISPR-Cas9 in plants. To enhance the utility of Cas12a, we investigate three Cas12a orthologs-Mb3Cas12a, PrCas12a, and HkCas12a-in plants. Protospacer adjacent motif (PAM) requirements, editing efficiencies, and editing profiles are compared in rice. Among these orthologs, Mb3Cas12a exhibits high editing efficiency at target sites with a simpler, relaxed TTV PAM which is less restrictive than the canonical TTTV PAM of LbCas12a and AsCas12a. To optimize Mb3Cas12a, we develop an efficient single transcription unit (STU) system by refining the linker between Mb3Cas12a and CRISPR RNA (crRNA), nuclear localization signal (NLS), and direct repeat (DR). This optimized system enables precise genome editing in rice, particularly for fine-tuning target gene expression by editing promoter regions. Further, we introduced Arginine (R) substitutions at Aspartic acid (D) 172, Asparagine (N) 573, and Lysine (K) 579 of Mb3Cas12a, creating two temperature-tolerant variants: Mb3Cas12a-R (D172R) and Mb3Cas12a-RRR (D172R/N573R/K579R). These variants demonstrate significantly improved editing efficiency at lower temperatures (22 °C and 28 °C) in rice cells, with Mb3Cas12a-RRR showing the best performance. We extend this approach by developing efficient Mb3Cas12a-RRR STU systems in maize and tomato, achieving biallelic mutants targeting single or multiple genes in T
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