ArticleGenome biology2023
Systematic optimization of Cas12a base editors in wheat and maize using the ITER platform.
Article in Genome biology, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 34 papers.
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Who cites it
34 citing papers in PubMed, 75 citations in OpenAlex.
- Enhancing CRISPR-Cas12a base editing in plants with LbCas12a variants and introns.Journal of integrative plant biology · 2026Article
- Optimizations of Cas12a- and Cas12i-based adenine base editors for efficient precision editing in the plant genome.Journal of integrative plant biology · 2026Article
- Optimized tRNA processing and TREX2-SpCas9 fusion enable high-efficiency multiplex genome editing in plants.Plant communications · 2026Article
- Wheat's Up with CRISPR-Cas-Current Advances, Obstacles and Perspectives.International journal of molecular sciences · 2026Review
- Advances in gene cloning and functional genomics approaches for wheat (Triticum aestivum L.) improvement.The plant genome · 2026Review
- Advances in Genome Editing for Plant Disease Resistance Breeding.Plants (Basel, Switzerland) · 2026Review
- Overcoming breeding barriers with genome editing in autopolyploid crops.BMC plant biology · 2026Review
- Engineering hypercompact IscB nucleases for efficient and versatile genome editing in rice.Genome biology · 2026Article
- Efficient genome editing by CRISPR-Cas12a in common wheat via shoot apical meristem delivery.Plant physiology · 2025Article
- Advances and Applications of Plant Base Editing Technologies.International journal of molecular sciences · 2025Review
- CRISPR/Cas genome editing in soybean: challenges and new insights to overcome existing bottlenecks.Journal of advanced research · 2025Review
- Synergistic optimization enhancing the precision and efficiency of cytosine base editors in poplar.Communications biology · 2025Article
- CRISPR-Cas applications in agriculture and plant research.Nature reviews. Molecular cell biology · 2025Review
- A comprehensive all-in-one CRISPR toolbox for large-scale screens in plants.The Plant cell · 2025Article
- Development of Protoplast-Based Gene Editing System for Areca Palm.Plants (Basel, Switzerland) · 2025Article
- Computationally derived RNA polymerase III promoters enable maize genome editing.Frontiers in plant science · 2025Article
- Expanding plant genome editing scope and profiles with CRISPR-FrCas9 systems targeting palindromic TA sites.Plant biotechnology journal · 2024Article
- An optimised CRISPR Cas9 and Cas12a mutagenesis toolkit for Barley and Wheat.Plant methods · 2024Article
- High performance TadA-8e derived cytosine and dual base editors with undetectable off-target effects in plants.Nature communications · 2024Article
- Expanding the targeting scope of CRISPR/Cas9-mediated genome editing by Cas9 variants inaBIOTECH · 2024Article
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Authors and funding
16 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundTesting an ever-increasing number of CRISPR components is challenging when developing new genome engineering tools. Plant biotechnology has few high-throughput options to perform iterative design-build-test-learn cycles of gene-editing reagents. To bridge this gap, we develop ITER (Iterative Testing of Editing Reagents) based on 96-well arrayed protoplast transfections and high-content imaging.
resultsWe validate ITER in wheat and maize protoplasts using Cas9 cytosine and adenine base editors (ABEs), allowing one optimization cycle - from design to results - within 3 weeks. Given that previous LbCas12a-ABEs have low or no activity in plants, we use ITER to develop an optimized LbCas12a-ABE. We show that sequential improvement of five components - NLS, crRNA, LbCas12a, adenine deaminase, and linker - leads to a remarkable increase in activity from almost undetectable levels to 40% on an extrachromosomal GFP reporter. We confirm the activity of LbCas12a-ABE at endogenous targets in protoplasts and obtain base-edited plants in up to 55% of stable wheat transformants and the edits are transmitted to T1 progeny. We leverage these improvements to develop a highly mutagenic LbCas12a nuclease and a LbCas12a-CBE demonstrating that the optimizations can be broadly applied to the Cas12a toolbox.
conclusionOur data show that ITER is a sensitive, versatile, and high-throughput platform that can be harnessed to accelerate the development of genome editing technologies in plants. We use ITER to create an efficient Cas12a-ABE by iteratively testing a large panel of vector components. ITER will likely be useful to create and optimize genome editing reagents in a wide range of plant species.
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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.