ArticlePlant biotechnology journal2021
Efficient gene targeting in Nicotiana tabacum using CRISPR/SaCas9 and temperature tolerant LbCas12a.
Article in Plant biotechnology journal, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 38 papers.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
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.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
38 citing papers in PubMed, 78 citations in OpenAlex.
- Wheat's Up with CRISPR-Cas-Current Advances, Obstacles and Perspectives.International journal of molecular sciences · 2026Review
- dSaCas9 enables enhanced transcriptional activation in Nicotiana benthamiana compared to its dSpCas9 ortholog.BMC plant biology · 2026Article
- Astaxanthin biofortification enhances tobacco tolerance to lead stress through boosting antioxidant defense, reducing Pb accumulation, and modulating detoxification pathways.Journal of advanced research · 2026Article
- Advances in the use of morphogenic regulators and peptide regenerating factors for boosting plant transformation and genome editing.Frontiers in plant science · 2026Review
- Casδ, an evolutionary transitional CRISPR system enables efficient genome editing across animals and plants.Nucleic acids research · 2025Article
- Application and Progress of Genomics in Deciphering the Genetic Regulation Mechanisms of Plant Secondary Metabolites.Plants (Basel, Switzerland) · 2025Review
- A comprehensive all-in-one CRISPR toolbox for large-scale screens in plants.The Plant cell · 2025Article
- Beyond a few bases: methods for large DNA insertion and gene targeting in plants.The Plant journal : for cell and molecular biology · 2025Review
- Enhancing CRISPR-Cas-based gene targeting in tomato using a dominant-negativeHorticulture research · 2025Article
- Effects of low temperature stress on photosynthetic characteristics and antioxidant defense system of flue-cured tobacco.Frontiers in plant science · 2025Article
- Versatile plant genome engineering using anti-CRISPR-Cas12a systems.Science China. Life sciences · 2024Article
- Context effects on repair of 5'-overhang DNA double-strand breaks induced by Cas12a in Arabidopsis.Plant direct · 2024Article
- Hydroponic Growth of [Journal of agricultural and food chemistry · 2024Article
- Cas12a-mediated gene targeting by sequential transformation strategy in Arabidopsis thaliana.BMC plant biology · 2024Article
- Determinants of CRISPR Cas12a nuclease activation by DNA and RNA targets.Nucleic acids research · 2024Article
- Cas12a and MAD7, genome editing tools for breeding.Breeding science · 2024Article
- Optimizing ErCas12a for efficient gene editing in Arabidopsis thaliana.Plant biotechnology journal · 2024Article
- Simple promotion of Cas9 and Cas12a expression improves gene targeting via an all-in-one strategy.Frontiers in plant science · 2024Article
- Recent advances in gene therapy: genetic bullets to the root of the problem.Clinical and experimental medicine · 2023Review
- Recent Trends and Advancements in CRISPR-Based Tools for Enhancing Resistance against Plant Pathogens.Plants (Basel, Switzerland) · 2023Review
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Nicotiana tabacum is a non-food herb that has the potential to be utilized as bio-factory for generating medicines, vaccines or valuable small metabolites. To achieve these goals, the improvement of genetic tools for pre-designed genome modifications is indispensable. The development of CRISPR/Cas nucleases allows the induction of site-specific double-strand breaks to enhance homologous recombination-mediated gene targeting (GT). However, the efficiency of GT is still a challenging obstacle for many crops including tobacco. Recently, studies in several plant species indicated that by replacing SpCas9 with other CRISPR/Cas-based nucleases, GT efficiencies might be enhanced considerably. Therefore, we tested SaCas9 as well as a temperature-insensitive version of LbCas12a (ttLbCas12a) for targeting the tobacco SuRB gene. At the same time, we also optimized the protocol for Agrobacterium-mediated tobacco transformation and tissue culture. In this way, we could improve GT efficiencies to up to a third of the inoculated cotyledons when using ttLbCas12a, which outperformed SaCas9 considerably. In addition, we could show that the conversion tract length of the GT reaction can be up to 606 bp long and in the majority of cases, it is longer than 250 bp. We obtained multiple heritable GT events, mostly heterozygous, but also biallelic GT events and some without T-DNA integration. Thus, we were not only able to obtain CRISPR/Cas-based heritable GT events in allotetraploid Nicotiana tabacum for the first time, but our results also indicate that ttLbCas12a might be a superior alternative for gene editing and GT in tobacco as well as in other crops.
Indexed as
Identifiers
What OpenQuestion holds
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.