Lentiviral Vector Design for Gene Silencing

Gene silencing by RNA interference (RNAi) has become a powerful and rapidly evolving method for investigating gene function in mammalian cells, especially small interfering RNA (siRNA) and short-hairpin RNA (shRNA). Lentiviral vectors (LVs) provide a unique tool to integrate these RNA expression cassettes with the goal of locally knocking down the expression of specific genes to assess the function of a gene. Creative Biolabs has proprietary technology and versatile platform in viral vector design, which enables us to provide excellent gene silencing LVs design services for our customers.

Introduction of Gene Silencing and LVs

RNAi, an RNA-mediated gene silencing mechanism, has recently emerged as a novel pathway that allows modulation of gene expression. As a tool in mammalian cell systems, gene silencing is induced by the delivery of a double-stranded RNA (dsRNA) that matches the mRNA target sequence. The dsRNA can be delivered as a siRNA via transfection, or shRNA via transfection or viral delivery of a plasmid. Because siRNA or shRNA can suppress the expression of genes of interest, RNAi-mediated gene silencing has become an essential technology to study gene functions in mammalian cells. To exploit this technique, efficient siRNA/shRNA delivery methods must be developed. Studies have found that LVs can be engineered to achieve stable and efficient gene silencing in a variety of cells. In gene-silencing LVs, siRNA can be delivered as a form of shRNA driven by RNA polymerase III promoters, or as a part of a miRNA-like structure expressed from RNA polymerase II promoter. Nowadays, gene-silencing LVs have been successfully used to inhibit HIV-1 infection, delay the occurrence and progression of amyotrophic lateral sclerosis in mice, prolong the survival rate of mice with scrapie development and increase the expression of fetal hemoglobin.

Application of Gene-Silencing LVs

Gene Silencing Lentiviral Vectors Design

Now RNAi-based silencing (e.g. siRNA and shRNA) is widely used for gene knockdown in basic scientific research and provides promise to new therapeutic strategies for a variety of diseases. Compared to knockout genetics, RNAi-based silencing is fast, cost-effective, and can be easily adapted to study homologous gene function in a wide variety of organisms. Furthermore, LVs provides a unique tool to integrate siRNA/shRNA expression cassettes with the aim to locally knockdown expression of a specific gene, enabling to assess the function of a gene. For instance, LVs with their high tropism for neurons offer one of the most attractive options for delivering shRNAs to the central nervous system (CNS). LVs-mediated delivery of shRNA leads to sustained knockdown of gene expression in the brain and spinal cord in vivo. Therefore, the LVs-mediated RNA silencing is a powerful aid to probe gene functions and gene therapy of diseases of the CNS. Additionally, this method has also been successfully used to functionally silence genes in primary mammalian cells, stem cells and in transgenic mice, resulting in persistent knockdown of gene expression.

Service

Using in-house developed protocols, Creative Biolabs provides unique gene silencing LVs design which allows efficient expression of any siRNA by transfection or lentiviral infection of target cells. The scheme is based on a unique convergent promoter design to increase the efficiency of target gene knockdown without the need for a hairpin loop structure commonly used for single promoter vectors. Our protocol combines the specificity of siRNA-mediated silencing cassettes with the versatility of LVs to stably transduce a wide range of cell types. The combination of the lentiviral and siRNA technologies provides a powerful tool to achieve long-term down-regulation of specific target genes both in vitro and in vivo.

Service Highlight

  • Safe-to-use LVs can deliver your gene into a wide range of cell lines
  • The best price with the best quality in its class
  • Fast turnaround time
  • Expert technical support, full confidentiality, and on-time delivery

Creative Biolabs is a leading commercial supplier of viral vector technologies, offering a full range of high-quality custom services for LVs design, including LVs design for targeting stem cell research, cellular reprogramming, immune modulation, gene editing. Our LVs can be used for gene function research, preclinical target validation, gene therapy & vaccine development for academic use or industrial R&D. Please feel free to contact us for more information.

For research use only. Not intended for any clinical use.