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| Home -> Products & Services -> Custom Plasmid Vector Construction Services -> Custom Construction Service of PiggyBac Transposon Expression Plasmid Vector | |||||||||||||||||||||||
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Custom Construction Service of PiggyBac Transposon Expression Plasmid Vector The PiggyBac transposon is a DNA transposable element derived from insect cells. It has been widely used in the field of gene delivery due to its high efficiency and stable gene integration capabilities. RGBiotech offers custom construction service of PiggyBac transposon expression plasmid vector. We can design a personalized vector solution according to your experimental objectives. If you are interested in our PiggyBac transposon expression vector construction service, please feel free to contact us at admin@rgbiotech.com for more information and technical support. Choose PiggyBac and experience a safe, efficient, and flexible gene delivery solution! Why Choose RGBiotech? 1. Diversity of Promoters: Provide a variety of promoters such as CMV, EF1a, and Tet-On, supporting constitutive or inducible expression.2. Choice of Reporter Genes: Markers such as GFP, Luciferase, and mCherry are convenient for real-time monitoring and screening. 3. Multiple Resistance Markers: Puromycin, neomycin, hygromycin, etc., are suitable for the stable screening of different cell types. 4. Delivery Content: Provide plasmid DNA and sequencing files with a complete construction report. 5. Sequence Verification: Confirm the accuracy of the inserted fragment, transposase gene, and regulatory elements through sequencing to ensure there are no mutations or frameshift errors. 6. Timeline and Price: Depending on the complexity of the construction, starts from 2 weeks, and the price is competitive and cost-effective. 7. If you have specific requirements (such as specific promoters, selection markers, etc.), you can further communicate to customize the solution. Read More 1. Discovery of Transposons Transposons, also known as "jumping genes", were first discovered by the Nobel laureate Barbara McClintock in the 1950s. They are movable DNA sequences. According to the transposition mechanism, they are divided into two categories: 1) Class I Transposons (Retrotransposons): Through the "copy-paste" mechanism, they are first transcribed into RNA and then reverse-transcribed into DNA, which is randomly integrated into the genome. 2) Class II Transposons (DNA Transposons): Through the "cut-paste" mechanism, the DNA fragments are directly cut and transferred by the transposase encoded by themselves.2. Characteristics of PiggyBac Transposon PiggyBac was initially isolated from insect baculovirus and is a representative member of DNA transposons. Its structure includes: 1) Terminal Inverted Repeats (TIR) at Both Ends: Sub-terminal repeat sequences of 13bp and 19bp are used for the recognition of transposase. 2) Transposase Coding Region: An open reading frame of 1.8kb encodes a transposase (PBase) consisting of 594 amino acids.3. PiggyBac Transposition Mechanism The PiggyBac transposon system consists of two components: 1) Transposon Vector: Contains the gene of interest, reporter gene, antibiotic resistance marker, and flanking PiggyBac terminal repeat sequences (TRs). 2) Transposase Expression Vector: Expresses PiggyBac transposase, which mediates the excision of the transposon from the vector and its integration into the host genome. After co-transfecting the transposon vector and transposase expression vector into host cells, the transposase recognizes the TRs and excises the transposon from the vector, subsequently inserting it into TTAA sites in the host genome to achieve stable expression of the foreign gene.PiggyBac achieves precise integration through the following steps: 1) The transposase recognizes and cuts the TIR sequences at both ends of the transposon, forming a 3'-OH terminal. 2) The 3'-OH attacks the TTAA site of the host genome, forming a hairpin structure and inserting it into a new position. 3) The host repair mechanism fills the gap and completes the integration, and the TTAA repeat sequence is retained on both sides of the integration site. 4. Features of PiggyBac Transposon 1) High Safety: It is a non-viral vector, avoiding the risk of viral infection.2) Easy to Operate: Directly transfected by plasmids, without complex viral packaging processes. 3) Large Vector Capacity: It can insert exogenous genes of 10-20kb, supporting the co-expression of multiple genes. 4) Predictable Integration Site: It prefers the TTAA sequence, reducing the random damage to host genes. 5) Reversibility: Precise excision can be achieved through re-transposition, facilitating subsequent experimental adjustments. 6) Efficient Gene Integration: The PiggyBac transposon system has a high-efficiency gene integration ability, which can significantly improve the insertion efficiency of exogenous genes into the host genome. It is especially suitable for the construction of stable cell lines. 7)Stable Genetic Expression: The integrated genes can be stably inherited to daughter cells, avoiding the problem of unstable expression caused by random integration of viral vectors or transient transfection. 8) Low Immunogenicity: With the characteristic of being a non-viral vector, it reduces the host immune response and is suitable for in vitro and in vivo experiments as well as research on gene therapy. 9) Flexible Custom Design: It supports the modular assembly of various elements (such as promoters, tags, selection markers, etc.) to meet personalized experimental needs. Wide Host Applicability: It can be applied to a variety of cell types, including mammalian, insect, and plant cells, with strong compatibility. 5. Applications of PiggyBac Transposon The PiggyBac transposon is widely used in gene function research, gene therapy, cell reprogramming, and other fields.1) Gene Function Research: Construct stable overexpression or knockout cell lines for the study of disease mechanisms. 2) Transgenic Animal Models: By microinjecting fertilized eggs, transgenic mice, zebrafish, etc. have been successfully prepared, and the efficiency can reach 45%. 3) Mutation Screening Tool: It has a bias towards inserting into genes, which is suitable for genome function annotation and the construction of mutation libraries. 4) Cell Reprogramming: Reprogramming somatic cells into induced pluripotent stem cells (iPSCs) for disease modeling and drug screening. 5) Gene Therapy: Delivering therapeutic genes to target cells for the treatment of genetic diseases, cancer, etc. 6. PiggyBac Transposon vs Viral Vectors Compared to traditional viral vectors, the PiggyBac transposon offers the following advantages.
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