Integration-deficient Lentiviral Vector Service
In recent years, gene therapy has developed rapidly in clinical applications, and new gene therapy techniques have become a key component in the treatment of various human diseases. Creative Biolabs is a leading provider of CRO services and has developed excellent lentiviral vector design platforms related to gene therapy. We provide a number of integration-deficient lentiviral vector construction services to meet every customer's requirement.
Introduction of Integration-deficient Lentiviral Vector
Currently, there are several possible ways to compromise the integration-deficient lentiviral vector (IDLV). The most effective method at present is to produce mutated virus strains. This can be easily accomplished by replacing the standard gag/pol packaging plasmid with an IN-mutant strain. Many studies have reported the examining the effects of mutating specific amino acids of IN to produce mutant strains with impaired integration in lentiviruses. Because IN is multifunctional, and involves virion morphogenesis, reverse transcription, PIC nuclear translocation, and integration, many of these mutations have an impact on several viral functions, not only in connection with integration but also on the reduction in the amounts of viral DNA (class II mutations). In general, class I mutations specifically affect IN function regarding DNA cleavage and integration, but result in normal levels of viral DNA, which are normally created by substituting any one of the three amino acids of the catalytic triad (D64, D116, and E152 for HIV-1 IN). Mutating or deleting the attachment (att) sites at the ends of the viral genome, involved in recognition by IN and cut by it, has also been shown to hamper provirus formation if performed on both U3 and U5 ends, but less efficiently than IN mutations.
Figure 1. Generation of integration-deficient lentiviral vector. (Wanisch, 2009)
The Service of Integration-deficient Lentiviral Vector
IDLVs can be produced by using integrase mutations that specifically prevent proviral integration, resulting in the generation of increased levels of circular vector episomes in transduced cells. These lentiviral episomes lack replication signals and are gradually lost by dilution in dividing cells, but are stable in stationary cells. Therefore, the services we can provide include the following: transient gene expression of IDLV in proliferating cells, stable expression in nondividing cells in vitro and in vivo, specific immune responses, RNA interference, homologous recombination (gene repair, knock-in and knock-out), site-specific recombination, and translocation.
Creative Biolabs has focused on the development of gene therapies for years, we whole-heartedly cooperate with you to accomplish our shared goals. We have accumulated scientific experience from the accomplished projects and are very proud of our high-quality lentiviral vector design services to ensure your requirements are met. If you are interested in our services, please contact us for more details.
Reference
- Wanisch, K.; et al. (2009). Integration-deficient lentiviral vectors: a slow coming of age. Molecular Therapy. 17(8): 1316-1332.
