The mitochondrial genomes of plants are larger and more complex than those of animals. Plant mitochondrial genomes can range in size from 200 to 2,500 kb, but there are not many genes encoded in them. The Arabidopsis thaliana mitochondrial genome, which is larger than the chloroplast genome, has just over 50 genes. Mitochondrial genomes are very susceptible to recombination, so plant mitochondrial genomes exist mainly as thread-like structures incorporating many ring and branching molecules. Many existing genome editing tools are unavailable due to limitations in delivery methods to the mitochondria. Reliable techniques for editing mitochondrial DNA are, therefore, one of the frontiers of genome engineering, and the world's best scientists have been working for years to make this a reality.
Lifeasible has been working in plant genome engineering for many years and has overcome considerable difficulties in developing a procedure for plant mitochondrial genome editing. We have the experience and expertise to provide a customized plant mitochondrial genome editing service to help you explore all aspects of your plant mitochondria.
We can edit plant mitochondrial genomes using transcriptional activator-like effector nucleases (TALENs) technology with mitochondrial localization signals (mitoTALENs). mitoTALENs technology uses a single protein to localize the mitochondrial genome, cleave the DNA into the desired gene and delete the target gene.
When using this technology for plant mitochondrial genome editing services, we broadly go through these steps: preparation of plant material to be edited; selection of target sequences; construction and transformation of plasmids, with transformed healing tissues cultured on kanamycin-containing medium; PCR and Southern blot analysis; extraction of total DNA from transgenic plants for NGS sequencing and short read coverage analysis, as well as genome structure analysis; and de novo sequencing to determine homologous recombination.
We can also provide you with plant mitochondrial genome editing services using a gene editing technology called Golden Gate. We first used the Golden Gate cloning system to construct the mt-DdCBE encoding plasmid for the target gene of the plant to be edited. Then, the plasmids are transformed into protoplasts of the plants to be edited. After a period of incubation in streptomycin or spectacular medium, the total DNA of the transgenic plants was extracted and verified by Sanger sequencing.
Due to the extreme complexity of the plant mitochondrial genome and the low density of coding genes, its research level is far less than that of the chloroplast genome. Therefore, the plant species currently capable of mitochondrial genome editing are minimal. We provide mitochondrial gene editing services, including but not limited to the following species.
Lifeasible has been exploring the frontiers of genome engineering since its inception. Our talented staff, extensive experience, and comprehensive technology are the basis for providing you with solutions for mitochondrial genome editing in plants. If you have an interest, questions, or collaboration, please contact us directly.
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Lifeasible has established a one-stop service platform for plants. In addition to obtaining customized solutions for plant genetic engineering, customers can also conduct follow-up analysis and research on plants through our analysis platform. The analytical services we provide include but are not limited to the following:
