Plant Gene Knockout Service
The CRISPR/Cas9 system has been widely used in many plant species to induce genomic mutations for researching gene function and improving crops. CD BioSciences has developed a CRISPR/Cas9-based plant knockout platform dedicated to enhancing excellent traits for engineered plants.
What is CRISPR/Cas9-Based Plant Gene Knockout
The sgRNA (short guide RNA) in the CRISPR/Cas9 system recognizes and binds to the target sequence of the target gene, which guides Cas9 to shear the binding site, producing a double-strand break (DSB). The organism repairs the DSB by non-homologous end joining (NHEJ), and the proteins involved in the repair insert or delete a few bases at the end of the DNA, thus realizing gene knockout in plants.
Figure 1. Flow diagram of the CRISPR/Cas9-based plant gene knockout. (Lawrenson, T., et al., 2019) Service Content
Service flow
- Design and synthesis of sgRNA.
- Vector construction and activity validation.
- Transformation.
- T0 generation culture.
- Genotype identification.
- Positive seedlings hardening and T1 seed production.
Comparison with second-generation gene editing technology (TALEN)
| Type | TALEN | CRISPR/Cas9 |
| Recognize target sequence patterns | Protein-DNA | RNA-DNA |
| Matching target sequence module | An RVD recognizes a single base | RNA-DNA base complementary pairing |
| Number of target sequence bases | >16bp*2 | About 20bp |
| Off-target rate | Low | Potentially off-target |
| Simultaneous knockout of multiple genes | No | Yes |
| Methylation sensitivity | Sensitive | Insensitive |
Other plant knockout services
- T-DNA insertion inactivation service
Our T-DNA insertion inactivation technology is widely used in plant gene knockout. The services include
- Agrobacterium Ti vector (tumor-inducing plasmid) construction.
- Transfection of Agrobacterium tumefaciens to form Agrobacterium tumefaciens.
- T-DNA import (requires the cooperation of multiple transposases and regulatory proteins).
- T-DNA integration.
- Formation of regenerative tissues in plants (healing tissues or regenerating plants).
- Conditional knockout service
In addition, we also achieve time- and space-specific gene knockouts by localized recombination systems. Our services include the Cre/Loxp and FLP/FRT systems.
- Cre/LoxP recombinase system
Our Cre/LoxP recombinase system allows DNA modifications to be made to specific cell populations or for the modification process to be triggered by specific external stimuli. It is ideal for researching complex cells in plants.
- FLP/FRT recombinase system
The yeast-derived FLP/FRT site-specific recombinase system allows us to research higher eukaryotic model organisms such as Arabidopsis thaliana and Oryza sativa due to its high recombination efficiency and targeting.
How to Place an Order?
At CD BioSciences, we offer comprehensive solutions for plant-targeted gene knockouts. With our cutting-edge facilities and experienced scientists, we can research gene function, unravel complex biological pathways, and accelerate progress in plant research. If you have any questions, please feel free to contact us. We are committed to delivering high-quality results and ensuring accuracy, efficiency, and confidentiality.
References
- Lawrenson, T., et al. (2019). Creating Targeted Gene Knockouts in Brassica oleracea Using CRISPR/Cas9. Methods in molecular biology (Clifton, N.J.). 1917, 155-170.
- Weeks, D. P., et al. (2016). Use of designer nucleases for targeted gene and genome editing in plants. Plant Biotechnology Journal. 14(2), 483-495.
For research use only, not for clinical use.