Solanum lycopersicum Transformation

Solanum lycopersicum or Lycopersicon esculentum Miller, also known as tomato, belong to the tomato family Solanaceae, and is an annual or perennial herbaceous plant. Solanum lycopersicum is rich in nutrients and has unique flavor, like carotene, lycopene and other nutrients, which are strong antioxidants that can effectively remove free radicals in the body, prevent and repair cell damage, and inhibit the spread and replication of cancer cells. Favored by consumers, it is now widely grown as an important edible fruit and vegetable in the world. In recent years, research on genetically modified Solanum lycopersicum has become a hot field to improve Solanum lycopersicum disease resistance, insect resistance, herbicide resistance, salt tolerance, cold tolerance, storage tolerance, fruit flavor, lycopene yield, etc.

Lifeasible provides one-stop services, covering all steps including experimental design, vector construction, plasmid transformation, positive transplant screening and characterization of transgenic Solanum lycopersicum. We offer Solanum lycopersicum transformation using various genetic engineering technologies as follows:

• Gene Overexpression

Through the quantitative overexpression of genes related to lycopene, anthocyanin, malic acid and other chemical components in Solanum lycopersicum, the production of specific compounds can be increased. We could help you overexpress many enzymes of the Rohmer pathway which might be critical for lycopene biosynthesis, SlMYB12 gene that has an important effect on the color of Solanum lycopersicum, and many other genes related to important traits of Solanum lycopersicum.

• RNA Interference (RNAi)

RNAi technology is widely used in the field of gene editing, it refers to the phenomenon of highly conserved, induced by double-stranded RNA (dsRNA), homologous mRNA high-efficiency and specific degradation in the evolutionary process. Through RNAi technology, the silencing of multiple genes in Solanum lycopersicum can be mediated.

• Virus-Induced Gene Silencing (VIGS)

Virus induced gene silencing (VIGS) means that after viruses carrying target gene fragments infect plants, they can induce plant endogenous gene silencing and cause phenotypic changes, and then study the function of target genes based on phenotypic variation. The VIGS technology is a method of transient transformation and underlying molecular basis may be post-transcriptional gene silencing. Silencing and functional analysis of target genes in Solanum lycopersicum through VIGS can help you save time and achieve valuable information for gene functional analysis. With wealth of experience in VIGS, the scientists in Lifeasible provides you customized protocol for VIGS in Solanum lycopersicum. We could achieve the transformation for Solanum lycopersicum with different genetic backgrounds.

• Gene Knockout with CRISPR Technology

The discovery of the CRISPR/Cas9 system provides us with a very powerful and convenient gene editing tool. By using CRISPR technology, we can knockout Solanum lycopersicum genes in different ways, including frameshift mutations, multiple deletion of fragments, knockout of non-coding genes, knockout of multiple copies of genes, etc.

• Gene Knock-in with CRISPR Technology

CRISPR system has strong scalability, and this scalability can be used to develop more useful gene editing tools. we have developed many methods that can improve gene knock-in efficiency and achieve precise editing of the Solanum lycopersicum genome. For the gene knock-in process, most of CRISPR gene knock-in is done through HDR. However, NHEJ and HDR will occur at the same time due to DNA breaks. Therefore, we have developed different methods to increase the probability of HDR, thereby improving the efficiency of gene knock-in.

• CRISPR Single Base Editing

CRISPR Single base editing technology is a hot area of life science research today. As a company that has been cultivating gene editing technology for decades, Lifeasible could help you achieve the conversion from C to T or A to G in Solanum lycopersicum using CBE and ABE, both of which rely on the DNA positioning capabilities of the CRISPR/Cas9 system. During single base editing, the C base deaminase or A base deaminase is located at a specific position in the genome, and it catalyzes the deamination reaction of C or A at a specific position and turns it into U or I. Then it is treated as T or G in the process of DNA replication, realizing the conversion from C to T or A to G.

• CRISPR Interference (CRISPRi)

As a variant of CAS protein after domain inactivation, dCAS9 still retains the ability to recognize and bind to specific DNA sequences, so it can derive many important functions. One of them is CRISPR Interference. There are many ways to participate in the inhibition of gene expression. For the inhibition of Solanum lycopersicum genes, we can provide a variety of solutions, including dCas9 binding to targeted DNA and realizing Inhibition of gene transcription through steric hindrance. In addition, gene knockdown can also be achieved by recruiting a fusion protein to the start site of gene transcription.

• CRISPR Activation (CRISPRa)

CRISPRa technology uses the powerful capabilities of Cas9 and sgRNA to fuse or recruit multiple proteins to enhance gene transcription. For Solanum lycopersicum genes, we provide VPR technology, SAM technology and Suntag technology to allow the CRISPR system to carry more activation element and achieve a stronger activation effect after synergistic amplification.

• CRISPR Mutation Library Construction

The study of gene function has always been the core subject of biological research. The earliest genetic screening system established through forward genetics is very inefficient and has a huge workload. However, the reverse genetic screening system based on CRISPR technology can complete very low-cost mutation library construction work. The gene mutation library construction technology we provide for Solanum lycopersicum including gene knockout library construction, gene knockdown library construction, and gene activation library construction. Moreover, single-cell sequencing is available for mutation screening.

• DNA-free Gene Editing

DNA-free gene editing technology has received extensive attention from the industry in recent years. We provide DNA free Solanum lycopersicum genome editing services, including transient expression of CRISPR/Cas9 plasmid DNA, in vitro transcription of CRISPR/Cas9, and pre-assembled ribonucleic acid composed of purified Cas9 protein and sgRNAs complex. These technologies can avoid the integration of foreign DNA and genome, and reduce off-target effects. In addition, compared with traditional techniques, these techniques can avoid the use of hybridization or backcrossing to isolate CRISPR/Cas9 chimeras, so they are cheaper and have shorter experimental cycles.

Genetic Transformation Process for Solanum lycopersicum

So far, the most advanced and widely used method for the development of genetically modified Solanum lycopersicum is Agrobacterium-mediated transformation. Briefly, the foreign target gene is transferred and integrated into plant cells through Agrobacterium, and then transformed plants are regenerated through techniques such as tissue culture.

Figure 1. Agrobacterium-mediated Solanum lycopersicum transformation by infecting cotyledon and hypocotyl with Agrobacterium (a) Regenerated kanamycin-resistant shoots from hypocotyl; (b) regenerated kanamycin-resistant shoots from cotyledon; (c) rooted kanamycin-resistant plantlet; (d) histochemical GUS staining of kanamycin-resistant plantlet leaf; (e) growth of transgenic lines in the greenhouse (Sun S, et al. 2015)

Lifeasible offers our customers with professional one-stop services, covering all steps including experimental design, vector construction, plasmid transformation, positive transplant screening and testing. Adapting to diverse purposes of different customers, multiple Agrobacterium strains (LBA288, NCPPB2695, GV3101, AG63, AGL-1, EHA105, and C58C1), as well as commercial and customized binary vectors with variant selectable markers (Kanamycin, Hygromycin, Phosphinothricin, G418, etc.) are readily available for your use. Experts at Lifeasible obtain comprehensive knowledge and years of experience to solve technical problems and challenges in Solanum lycopersicum transformation. We can draw customized solution to help you research on a variety of Solanum lycopersicum genes (Including SGR1, LCY-E, Blc, LCY-B1 and LCY-B2 that play a key role in Carotenoid metabolism, and GABA-TP1, TP2, TP3, GAD2, GAD3 and ALMT9 gene family and other genes that have a significant impact on Solanum lycopersicum GABA metabolic pathway and Malic acid synthesis pathway, etc.), Our services guarantee the success of your project. For more information or any inquiry requirements, please contact Lifeasible.