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Pseudotyping


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Pseudotyping

Pseudotyping is the process of producing viruses or viral vectors in combination with foreign viral envelope proteins. Therefore, a pseudotyped virus (PV) is one in which the outer shell (the envelope glycoproteins of an enveloped virus or the capsid proteins of a nonenveloped virus) originates from a virus that differs from the source of the genome and the genome replication apparatus. A PV is capable only of entering and transducing a target cell, initiating nucleic acid replication without producing infectious particles.

Usually, a functional PV can be generated by transfection of a “producer” cell with three plasmids. Using the production of pseudotyped retroviruses as an example, the three plasmids include: i) One contains the gag and pol genes that encode the retroviral core structural proteins and replication enzymes; ii) One contains the viral long terminal repeats (LTRs), which possess the signals for transcription and integration of the viral genome, the Ψ RNA packaging signal, which promotes the specific encapsidation of the transcribed RNA into recombinant particles, and the gene that is transduced; iii) One expresses the open reading frame of the virus envelop protein (VEP) of the virus of interest with appropriate signal peptide.

pseudotyping process for pseudotyped virus.

Figure 1 Schematic of the basic three-plasmid approach to generation of a PV.
(Future Virology 2016)

Pseudotyping Services at Creative Biogene

Creative Biogene has accumulated plenty of experience working with pseudotyping. With strict consideration of different factors, including viral backbones for pseudotyping, provision of proteases, signal peptides and transmembrane domains, and VEP alteration, we offer pseudotyped viruses for a wide range of applications.  

Creative Biogene has established a standardized and simplified PV production platform with low cost, with optimization of quantification of PVs, concentration of PVs, influence of plasmid and codon, and the use of PV packaging cell lines. We provide commonly used vectors in gene therapy trials, such as VSV G-pseudotyped lentivirus.

Applications of PVs:

  • Research and therapeutic agent screening
  • Measuring antibodies
  • Vaccines
  • Gene transfer

References:

  1. Sanders, D.A. (2002) No false start for novel pseudotyped vectors. Current Opinion in Biotechnology. 13(5): 437-442.
  2. King, B., Temperton, N.J., Grehan, K., Scott, S.D., Wright, E., Tarr, A.W., Daly, J.M. (2016) Technical considerations for the generation of novel pseudotyped viruses. Future Virology. 11(1): 47-59.

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