ArticlePlant biotechnology journal2019
Single transcript unit CRISPR 2.0 systems for robust Cas9 and Cas12a mediated plant genome editing.
Article in Plant biotechnology journal, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 89 papers.
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Who cites it
89 citing papers in PubMed, 153 citations in OpenAlex.
- Establishment of an efficient and PAM-relaxed LbCas12a genome editing tool in plants.The New phytologist · 2026Article
- The plant Golden Gate toolkit: compatibility, interoperability, and harmonized assembly in plant synthetic biology.Plant methods · 2026Review
- Protein post-translational modifications: key switches coordinating fruit ripening regulatory networks.Horticulture research · 2026Article
- Transgene-free genome editing in citrus and poplar trees using positive and negative selection markers.Plant cell reports · 2025Article
- Programmable genome engineering and gene modifications for plant biodesign.Plant communications · 2025Review
- Evolutionary and Structural Analysis of the Aquaporin Gene Family in Rice.Plants (Basel, Switzerland) · 2025Review
- Harnessing promoter elements to enhance gene editing in plants: perspectives and advances.Plant biotechnology journal · 2025Review
- Harnessing novel cytidine deaminases from the animal kingdom for robust multiplexed base editing in rice.Plant biotechnology journal · 2025Article
- CRISPR-Cas12i confers efficient genome editing and gene regulation in plants.Plant physiology · 2025Article
- Review
- Engineering a bacterial toxin deaminase from the DYW-family into a novel cytosine base editor for plants and mammalian cells.Genome biology · 2025Article
- An efficient CRISPR-Cas12a-mediated MicroRNA knockout strategy in plants.Plant biotechnology journal · 2025Article
- PAM-relaxed and temperature-tolerant CRISPR-Mb3Cas12a single transcript unit systems for efficient singular and multiplexed genome editing in rice, maize, and tomato.Plant biotechnology journal · 2025Article
- Improving rice grain shape through upstream ORF editing-mediated translation regulation.Plant physiology · 2024Article
- Identification of multiple miRNA-encoded peptide reveals OsmiPEP162a putatively stabilizes OsMIR162 in rice.Plant cell reports · 2024Article
- Versatile plant genome engineering using anti-CRISPR-Cas12a systems.Science China. Life sciences · 2024Article
- IsDge10 is a hypercompact TnpB nuclease that confers efficient genome editing in rice.Plant communications · 2024Article
- Advancing PAM-less genome editing in soybean using CRISPR-SpRY.Horticulture research · 2024Article
- High performance TadA-8e derived cytosine and dual base editors with undetectable off-target effects in plants.Nature communications · 2024Article
- Boosting genome editing in plants with single transcript unit surrogate reporter systems.Plant communications · 2024Article
29 more citing papers are in PubMed but not listed here.
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Authors and funding
16 authors at 4 institutions in 3 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
CRISPR-Cas9 and Cas12a are two powerful genome editing systems. Expression of CRISPR in plants is typically achieved with a mixed dual promoter system, in which Cas protein is expressed by a Pol II promoter and a guide RNA is expressed by a species-specific Pol III promoter such as U6 or U3. To achieve coordinated expression and compact vector packaging, it is desirable to express both CRISPR components under a single Pol II promoter. Previously, we demonstrated a first-generation single transcript unit (STU)-Cas9 system, STU-Cas9-RZ, which is based on hammerhead ribozyme for processing single guide RNAs (sgRNAs). In this study, we developed two new STU-Cas9 systems and one STU-Cas12a system for applications in plants, collectively called the STU CRISPR 2.0 systems. We demonstrated these systems for genome editing in rice with both transient expression and stable transgenesis. The two STU-Cas9 2.0 systems process the sgRNAs with Csy4 ribonuclease and endogenous tRNA processing system respectively. Both STU-Cas9-Csy4 and STU-Cas9-tRNA systems showed more robust genome editing efficiencies than our first-generation STU-Cas9-RZ system and the conventional mixed dual promoter system. We further applied the STU-Cas9-tRNA system to compare two C to T base editing systems based on rAPOBEC1 and PmCDA1 cytidine deaminases. The results suggest STU-based PmCDA1 base editor system is highly efficient in rice. The STU-Cas12a system, based on Cas12a' self-processing of a CRISPR RNA (crRNA) array, was also developed and demonstrated for expression of a single crRNA and four crRNAs. Altogether, our STU CRISPR 2.0 systems further expanded the CRISPR toolbox for plant genome editing and other applications.
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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.