Gene editing is a technology that allows for targeted modification of the genome and is the basis for studying the function of specific genes. In recent years, the main genome editing technologies are zinc finger nuclease (ZFN) and transcription activator-like effector nucleases (TALEN). The CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins) system, as a new generation of genome editing technology, has received a lot of attention and great response from researchers since its introduction.
Figure 1. The mechanism of RNA guided DNA cleavage mediated by Cas9 (Khaoula et al. 2013)
Currently, the CRISPR/Cas9 system-mediated third-generation genome editing technology has been successfully applied to genome modification in many species, including model and crop plants. There are many strategies for genes knockout. Among them, the least impact on the cell should be to cause frameshift mutations in the coding sequence of the gene. Use Cas9/sgRNA to act on the front exons of the gene to cause random mutations upstream of the coding sequence of the gene, it is easy to cause frameshift mutation. Genes with frameshift mutations generally have two results: the downstream coding sequence is likely to have stop codons in advance, triggering nonsense-mediated mRNA decay (NMD), the mRNA signal of the gene cannot be detected; even if there is no premature termination, the amino acid sequence encoded by the downstream sequence after the frameshift mutation has been completely changed, and the translated protein generally loses its function. No matter what happens, on the basis of minimal changes to the genome, the amino acid sequence encoded by the gene is changed significantly, the function of the gene is destroyed, and the specific gene is knocked out.
Fig 2. Gene knockout experiment procedure (Khaoula et al. 2013)
Lifeasible is a world-class phytotechnology company with extensive experience in plant gene knockout using CRISPR/CAS9 technology. We provide a complete knockout solution for you, from knockout protocol design→knockout site design→vector construction→transformation→knockout assay→result analysis, so that your experiment is successful in one go. We provide the most cost-effective service, efficient and professional is what our customers say most about Lifeasible. Not only you can save money on your project by choosing Lifeasible, we also guarantee absolute data security and will never disclose your experiments and related results to anyone without your consent.
We have more than 5 years of rich operational experience in the field of genome editing and have been engaged in the research and development of plant genome editing technology for a long time.
We provide one stop service from sgRNA design and vector construction, plant transformation and screening, mutation identification, RNA and protein level identification, even phenotype analysis to seed harvest. We have established fully-fledged gene editing platform in many species such as Arabidopsis, rice, wheat, alfalfa, corn, cotton, sorghum, soybean, sugarbeet, cabbage, tobacco, eggplant, cucumber, potato, petunia, oilseed rape, and so on.
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Lifeasible offers complete, professional plant gene knockout solutions, as well as customized experimental protocols based on your project requirements and plant sample characteristics. For more information, please contact Lifeasible.