ArticleThe New phytologist2023
Combining multiplex gene editing and doubled haploid technology in maize.
Article in The New phytologist, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
17 citing papers in PubMed, 21 citations in OpenAlex.
- Advances and Applications of Genome Editing in Precision Crop Breeding.Plants (Basel, Switzerland) · 2026Review
- Engineering ultra-low-gliadin wheat for celiac disease using an integrated RNAi, CRISPR, and doubled haploid strategy.Journal of experimental botany · 2026Article
- Diverse molecular mechanisms of in vivo haploid induction in plants: from fertilization interference to genome programming.Plant cell reports · 2026Review
- Molecular Regulators of In Vitro Regeneration in Wheat: Roles of Morphogenic Factors in Transformation, Genome Editing, and Breeding.International journal of molecular sciences · 2026Review
- Genome editing beyond transgenes: DNA-free technologies in crop improvement.Frontiers in plant science · 2026Review
- Beyond the Edit: Integrating CRISPR-Cas Genome Editing Into the Cereal Breeding Pipeline.International journal of genomics · 2026Review
- Unlocking grass leaf development: foundations for tunable cereal design.The New phytologist · 2025Review
- Turbo-charging crop improvement: harnessing multiplex editing for polygenic trait engineering and beyond.The Plant journal : for cell and molecular biology · 2025Review
- Harnessing haploid-inducer mediated genome editing for accelerated maize variety development.Plant biotechnology journal · 2025Article
- In vitro- and in vivo-based approaches for doubled haploid production in Zea mays L.: challenges and opportunities.TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik · 2025Review
- Genetic Analysis and Construction of a Fingerprint for LicensedPlants (Basel, Switzerland) · 2024Article
- Article
- Breeding for improved digestibility and processing of lignocellulosic biomass inFrontiers in plant science · 2024Review
- Roles of auxin pathways in maize biology.Journal of experimental botany · 2023Review
- Unlocking Nature's Clock: CRISPR Technology in Flowering Time Engineering.Plants (Basel, Switzerland) · 2023Review
- From Petri Dish to Field: Plant Tissue Culture and Genetic Engineering of Oats for Improved Agricultural Outcomes.Plants (Basel, Switzerland) · 2023Review
- Green revolution to genome revolution: driving better resilient crops against environmental instability.Frontiers in genetics · 2023Review
Corrections and comments
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Authors and funding
12 authors at 1 institution in 1 country.
Funding
Abstract
A major advantage of using CRISPR/Cas9 for gene editing is multiplexing, that is, the simultaneous targeting of many genes. However, primary transformants typically contain hetero-allelic mutations or are genetic mosaic, while genetically stable lines that are homozygous are desired for functional analysis. Currently, a dedicated and labor-intensive effort is required to obtain such higher-order mutants through several generations of genetic crosses and genotyping. We describe the design and validation of a rapid and efficient strategy to produce lines of genetically identical plants carrying various combinations of homozygous edits, suitable for replicated analysis of phenotypical differences. This approach was achieved by combining highly multiplex gene editing in Zea mays (maize) with in vivo haploid induction and efficient in vitro generation of doubled haploid plants using embryo rescue doubling. By combining three CRISPR/Cas9 constructs that target in total 36 genes potentially involved in leaf growth, we generated an array of homozygous lines with various combinations of edits within three generations. Several genotypes show a reproducible 10% increase in leaf size, including a septuple mutant combination. We anticipate that our strategy will facilitate the study of gene families via multiplex CRISPR mutagenesis and the identification of allele combinations to improve quantitative 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.