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| Home -> Products & Services -> Custom Plasmid Vector Construction Services -> Custom Construction Service of Retroviral Expression Plasmid Vector (MMLV/MSCV) | ||
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Custom Construction Service of Retroviral Expression Plasmid Vector (MMLV/MSCV) Retroviral vectors are gene delivery tools derived from modified retroviruses, such as Moloney Murine Leukemia Virus (MMLV) and Murine Stem Cell Virus (MSCV). They enable stable integration of exogenous genes into the host genome, making them ideal for long-term gene expression in dividing cells. Engineered for safety and efficiency, retroviral vectors are widely used in gene therapy, cell reprogramming, and functional genomics. RGBiotech is committed to providing customers with professional, efficient, and high-quality retroviral plasmid vector construction services to contribute to the development of scientific research and the biomedical field. If you have any needs, please feel free to contact us at admin@rgbiotech. Why Choose RGBiotech? 1) Diverse Retroviral Backbone Plasmid Vectors: We have a rich library of empty vectors to meet different needs of our customers.2) Multiple Promoter Options: We provide a variety of promoters such as LTR, CMV, EF1α, U6 to precisely regulate gene expression. 3) Abundant Reporter Genes: We have reporter genes such as GFP, RFP, LacZ and Luciferase etc., which are convenient for monitoring the transduction efficiency of retrovirus. 4) Multiple Resistance Selection Markers: We have a complete range of resistance genes such as Neo, Hygro, and Puro to assist in the screening of stable cell lines. 5) We also provide the retroviral packaging plasmid vectors, which can be used to package and produce retroviral particles with different types of envelope proteins to optimize cell targeting and transduction efficiency. In addition to the common VSVG envelope protein, our packaging plasmids also include: a) Amphotropic Envelope Protein: It has a wide host range and can infect a variety of mammalian cells. b) Ecootropic Envelope Protein: It mainly infects mouse and rat cells and has a high infection efficiency for rodent cells. c) 10A1: Broadens tropism for specific rodent and human cells. d) RD114: Enhances infection in hematopoietic and primary cells. 6) Comprehensive Quality Control: All plasmid vectors are validated by sequencing to ensure 100% accuracy. 7) Cost Efficiency: Competitive pricing with bulk order discounts. Read More Retroviruses are a class of RNA viruses. Their genomes can be converted into DNA through the reverse transcription process and integrated into the host cell genome. Retroviral vectors are constructed using this characteristic to introduce foreign genes into host cells and achieve stable expression. This vector system usually consists of packaging plasmids and vector plasmids. The packaging plasmids provide the proteins required for virus packaging, and the expression or transfer plasmid vectors carry the target genes. 1. Applications of Retrovirus 1) Cell Biology Research: To stably transfect cell lines for studying gene functions, signal pathways, etc.a) Stable Cell Line Construction: Efficient establishment of mammalian cell lines with stable gene expression. b) Cell Reprogramming: Generation of induced pluripotent stem cells (iPSCs) or differentiated cell types. 2) Cancer Research: To construct cancer cell models for studying the mechanisms of cancer development and screening anti-cancer drugs. 3) Gene Therapy: Delivery of normal genes or therapeutic genes (e.g., cytokine genes, tumor suppressors) into patient cells to correct gene defects and treat genetic diseases. 2. Features of Retrovirus 1) Efficient Integration: Can efficiently integrate foreign genes into the host cell genome to achieve stable expression.2) Broad Host Range: Can infect various types of mammalian cells. 3) Low Immunogenicity: Compared with other viral vectors, it causes a weaker immune response. 3. Classification of Retrovirus: MMLV & MSCV 1) MMLV (Moloney Murine Leukemia Virus): It is one of the most commonly used retroviral vectors. Its advantages are simple construction and convenient operation. However, it has a preference for dividing cells and is difficult to infect non-dividing cells.2) MSCV (Murine Stem Cell Virus): Can infect a variety of stem cells, including hematopoietic stem cells, and can stably express foreign genes in stem cells, which is of great significance for stem cell-related research. 4. Differences between Retrovirus & Lentivirus 1) Infected Cell Types: Retroviruses mainly infect dividing cells, while lentiviruses can infect both dividing and non-dividing cells.2) Integration Methods: Retroviruses randomly integrate into the host genome, which may lead to insertional mutations; the integration sites of lentiviruses are relatively more inclined to transcriptionally active regions, with higher safety. 5. Elements Required for Retroviral Expression Plasmid Vectors 1) LTRs (Long Terminal Repeats): Drive proviral integration and gene expression.2) Ψ (Psi) Packaging Signal: Ensures viral RNA encapsidation. 3) Multiple Cloning Site (MCS): Flexible insertion of your gene of interest. 4) Safety Features: Self-inactivating (SIN) LTRs available to minimize insertional mutagenesis risks. 5) Promoters: Such as the LTR, CMV (Cytomegalovirus Promoter), EF1α (Elongation Factor 1α Promoter), etc., which drive the expression of foreign genes. 6) Reporter Genes: Such as GFP (Green Fluorescent Protein), RFP (Red Fluorescent Protein), Luciferase, etc., which are used to monitor virus transduction and gene expression. 7) Mammalian Resistance Selection Markers: Such as Neo (Neomycin Resistance Gene), Hygro (Hygromycin Resistance Gene), Puro (Puromycin Resistance Gene), etc., which are convenient for screening stably transfected cells. 6. Packaging Limit of Retrovirus The size of foreign fragments packaged by retroviral vectors generally does not exceed 8kb. Excessively large fragments may affect the virus packaging efficiency and infection ability.7. Retrovirus Packaging Process 1) Plasmid Vector Construction: Synthesize and clone the target gene into the retroviral expression plasmid vector.2) Co-transfection: Co-transfect the constructed expression plasmid vector and the packaging plasmids into the packaging cell line (such as HEK293T cells). 3) Virus Harvest: At a certain time after transfection (e.g., 48h), collect the cell culture supernatant containing the retroviral particles. 4) Virus Concentration: Concentrate the virus supernatant by PEG and/or ultracentrifugation to increase the retrovirus titer. 5) Virus Titer Determination: Use appropriate methods (such as fluorescence quantitative PCR, TCID50, etc.) to determine the virus titer to ensure the quality of the virus. |
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