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Simple lentivirus enrichment Everyone can get started easily

2022-09-23

Simple lentivirus enrichment Everyone can get started easily
Lentiviruses have the characteristics of high transfection efficiency, can infect a wide range of host cells (such as primary cells), carry larger gene fragments, target cell-specific activation (different promoters), etc., and are often used as the delivery of specific genes, cells Model building or disease research tools, by integrating genes into the host cell genome, can be combined with RNAi nucleic acid interference or CRISPR/Cas9 gene editing technology to perform gene expression (Gene expression), gene silence (Gene silence) and Gene editing, etc.

The use of lentivirus has been widely used in various fields. Whether it is academic research or clinical treatment, many related studies have been published. For example, the popular sars-cov-2 study, a research team established lentivirus-ACE2 model that can infect mice with lentivirus, so that human SARS-COV-2 virus can infect mouse lung epithelial cells, which is the research for the development of animal COVID-19 vaccine. In addition, some research teams use lentivirus as a marker of tumor cells, and lentivirus vector selects the polymerase (encoding Pol2) and specific promoter (FERH promoter) that can be expressed only in tumor to start the expression of fluorescent (GFP) protein, which can be used for the tracking and separation of tumor cells.

Lentivirus packaging system
Lentivirus can form a non-pathogenic pseudovirus by lentivector carrying target gene and packaging system, and embed interested siRNA, shRNA, miRNA, etc. into host genes for long-term expression. Currently, the most common lentivirus packaging belongs to the 4th generation, which is mainly composed of four plasmids:
  1. Packaging plasmid: mainly for the generation of the structure of the virus, including the gag /pol gene, the gag gene will make the outer sheath protein of the virus; the pol gene will make the replication enzyme of the virus
  2. Envelope plasmid: VSV-G gene that forms the viral envelope
  3. REV plasmid : The rev gene that forms a regulatory protein that regulates expression such as RNA splicing and protein transport
  4. Transfer plasmid: contains the target gene sequence, 5'LTR with a heterologous promoter (RSV), can insert the target sequence into the host gene
The 4th generation lentiviral vector system greatly improves safety and reduces the risk of forming replication-competent retroviruses.
 

How to efficiently separate and concentrate virus particles
TOOLSilent LentiPrecipitating Reagent is a mixture of polymers optimized for the precipitation of lentiviral particles. It provides a simple, fast, and highly efficient method for concentrating lentiviral particles.

Product features:
  1. Easy to use: simply mix.
  2. No ultracentrifugation required.
  3. Easily scale up for large volumes.
  4. Up to a 100-fold increase in concentration.
  5. Cost-effective spin protocol for efficient viral concentration.
  6. Nontoxic: safe for all cell lines, including ES cells.

Biotools provides complete lentivirus culture and isolation products

 
Product specification:

Reference:
  1. Jonghyuck Park, Joseph T. Decker, Dominique R. Smith, Brian J. Cummings, Aileen J. Anderson, Lonnie D. Shea , Reducing inflammation through delivery of lentivirus encoding for anti-inflammatory cytokines attenuates neuropathic pain after spinal cord injury. Corel (2018), doi:10.1016/j.jconrel.2018.10.003
  2. A. Lamsfus-Calle, A. Daniel-Moreno, G. Ureña-Bailén, et al., Hematopoietic stem cell gene therapy: The optimal use of Lentivirus and gene editing approaches, Blood Reviews(2019), doi:10.1016/j.blre.2019.100641
  3. Chi-Ping Day, John Carter, Carrie Bonomi, Dominic Esposito, Bruce Crise, Betty Ortiz-Conde, Melinda Hollingshead, and Glenn Merlino, Lentivirus-mediated bifunctional cell labeling for in vivo melanoma study, Pigment Cell Melanoma Res. (2009),  doi: 10.1111/j.1755-148X.2009.00545.x
 
 
 
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