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Lentiviruses efficiently deliver genetic information, ensuring long-term gene expression. Widely utilized in diverse mammalian cell types, lentiviral vectors are essential for stable transgene expression, gene silencing, immunization, transgenic animals, stem cell modifications, and more. Their ability to infect both dividing and non-dividing cells makes them particularly appealing for human gene therapy, supported by advancements in the recombinant lentivirus system.
As a leader in lentiviral technology, Creative Biogene has developed a comprehensive library of human, mouse, and rat genes cloned into lentiviral vectors or ready-to-use lentivirus, which can be used to manipulate the expression of your gene of interest within a wide range of host cells. Our comprehensive suite of lentiviral solutions includes ready-to-use particles suitable for use in cell culture and animals, plasmid clones expressing ORF, miRNA, shRNA, and sgRNA. Additionally, we offer lentivirus expression plasmid with desired promoters or tags.
Advantages of Our Premade Lentivirus Particles
Creative Biogene provides lentiviruses containing reporter genes such as GFP, RFP, luciferase etc.
Creative Biogene offers multiple lentiviral particles that can be used as control.
Creative Biogene has generated premade lentiviruses that can be used in the development of iPS cells.
Creative Biogene has developed ready-to-use lentiviruses that are useful tools in CAR-T cell generation.
Creative Biogene is offering recombinase lentiviruses that can be used in genome editing.
Creative Biogene offers lentiviral particles that can be used for studying exosomes.
Creative Biogene provides lentiviral particles containing various ORFs that can mediate target gene expression in host cells.
Creative Biogene offers lentiviral particles to support miRNA research.
Creative Biogene offers lentiviral particles that can be used for developing immortalized cell line.
Lentiviruses are a subclass of retroviruses with single-stranded RNA genomes that can integrate into host chromosomes. Lentiviral vectors have the parental virus genome split into plasmids containing packaging signals, key viral elements, and a gene transfer cassette. This design minimizes the chances of replication-competent virus generation while keeping integration capacity. Lentiviral vectors can efficiently integrate into the genomic DNA of dividing and non-dividing cells, offering advantages in transduction breadth, expression sustainability, and large cargo size. Hence, lentiviral vectors have broad utility for stable gene overexpression/knockdown, lineage tracing, vaccine engineering, and cell reprogramming applications.
Additionally, some key molecular biology applications of the listed lentiviral tools are:
Case Study 1
The tumor microenvironment, critical in various cancer processes, harbors chemotactic factors influencing tumorigenesis, inflammation, and therapeutic resistance. CXCL5, a key chemotactic factor, overexpressed in cancers, propels tumor proliferation, invasion, angiogenesis, and therapeutic resistance. Researchers reveal a significant correlation between the elevated expression of CXCL5 and multiple oncogenic signaling pathways in glioblastoma multiforme (GBM), including JAK-STAT and NF-κB signaling pathways. Furthermore, CXCL5 plays a crucial role in tumorigenesis and angiogenesis, highlighting the potential for novel therapeutic approaches targeting CXCL5 in GBM.
Figure 1. CXCL5 promotes cell proliferation, tumorigenesis, and angiogenesis in GBM. (Systematically introducing specific genes into cells was accomplished by utilizing lentiviral particles (LV10093L and LV00926Z) from Creative Biogene, NY.)(Mao P, et al., 2023)
Case study 2
Human intestinal tissue-derived steroids (HIEs) offer a powerful ex vivo model for gastrointestinal research. Using a CRISPR-Cas9 system with lentiviral transduction, researchers can efficiently knock out genes in HIEs. This protocol, distinct from electroporation methods, utilizes a modified LentiCRISPRv2-small-guiding RNA. It includes gene delivery, single-cell cloning, clone verification, and mutation site sequencing. The researcher is guided through a concise, efficient process, allowing the generation of a genetic knockout HIE line within 2-3 months. Additionally, HIEs serve as an effective model for studying host restriction factors for viral replication, as demonstrated with human norovirus. The use of lentiviral transduction ensures swift and precise genetic modifications in HIEs.
Figure 2. The lentivirus packaging and transduction protocol involves seven key steps: preparing a CRISPR-Cas9 lentivirus vector, packaging the lentivirus, transducing HIEs, recovering and selecting HIEs, performing single-cell cloning, sequencing verifying clones, and utilizing knockout lines to assess host gene function in viral replication or other biological processes (Lin, S. C., et al., 2022).
Q: What are lentiviruses?
A: Lentiviruses are a subgroup of retroviruses that can integrate into the host genome and achieve long-term transgene expression in both dividing and non-dividing cells. Lentiviral vectors are engineered from viruses like HIV-1 by removing all viral coding sequences and replacing them with a gene of interest.
Q: What steps are required to generate stable cell lines using lentiviral vectors?
A: The major steps include:
(1) Clone gene of interest into a lentiviral transfer vector
(2) Cotransfect packaging plasmids and transfer vector into 293T cells
(3) Collect and concentrate lentiviral particles
(4) Transduce target cells with lentiviral particles
(5) Select stably transduced cells using antibiotics or fluorescence-activated cell sorting
(6) Expand and validate transgene expression
Q: How are lentiviral packaging sequences designed?
A: Lentiviral packaging systems split essential viral components across several plasmids to maximize biosafety. Key elements are:
(1) Packaging plasmid: Encode structural and replication proteins
(2) Envelope plasmid: Encode the heterologous envelope protein like VSV-G
(3) Additional plasmids: Regulatory proteins like Rev
(4) Transfer vector: Contain cis-acting sequences for packaging and integration
Q: Do cell morphologies change after lentiviral transduction?
A: Most cell morphologies remain unaffected after lentiviral transduction if optimal transduction conditions are used. However, very high viral doses can lead to cellular toxicity and morphology changes. It's important to titer lentiviruses and use appropriate multiplicity of infection.
Q: How to determine lentiviral titer and assess transduction efficiency?
A: Methods to titer lentiviruses include:
(1) Measure RNA copy number by RT-qPCR
(2) Reporter-based flow cytometry by expressing fluorescent marker
(3) Measure integrated viral DNA copies per cell genome by qPCR
(4) Assess functional transgene expression by PCR, RT-qPCR, western blot, etc.
| Cat.No. | Product Name | Price |
|---|---|---|
| LV00014Z | Human NGFR lentiviral particles | Inquiry |
| LV00060Z | Human BCL6 lentiviral particles | Inquiry |
| LV00061Z | Human BRF1 lentiviral particles | Inquiry |
| LV00062Z | Human CAB39 lentiviral particles | Inquiry |
| LV00063Z | Human CEBPA lentiviral particles | Inquiry |
| LV00064Z | Human CEBPB lentiviral particles | Inquiry |
| LV00065Z | Human CEBPD lentiviral particles | Inquiry |
| LV00066Z | Human CREBBP lentiviral particles | Inquiry |
| LV00067Z | Human FOXO3 lentiviral particles | Inquiry |
| LV00068Z | Mus musculus Mmp3 lentiviral particles | Inquiry |
| LV00069Z | Human PRDM1 lentiviral particles | Inquiry |
| LV00071Z | Human PRKAA2 lentiviral particles | Inquiry |
| LV00073Z | Human SIRT1 lentiviral particles | Inquiry |
| LV00074Z | Mus musculus Sirt1 lentiviral particles | Inquiry |
| LV00075Z | Human SIRT2 lentiviral particles | Inquiry |
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