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GenomONE™ - Neo EX HVJ Envelope Transfection Kit

Background:

HVJ (hemagglutinating virus of Japan) Envelope VECTOR KIT is a tool for transfection of molecules (plasmid DNAs, siRNAs, oligonucleotides, proteins, antibodies etc.) into cells and animal tissue  by means of membrane fusion.
The HVJ envelope, carrying the molecule to be transfected, is composed of a completely inactivated and purified HVJ (Sendai virus) while preserving the cell membrane-fusing capability of the envelope.
GenomONE provides ready-to-use kits containing the HVJ envelope and auxiliary reagents (incorporation enhancer, incorporation reagent, introduction enhancer, buffer).

Principe:

What is HVJ Envelope (HVJ-E)?
HVJ Envelope is a purified product prepared through complete inactivation of Sendai virus (HVJ*). It is a vesicle in which only the cell membrane-fusing capability of the envelope protein of Sendai virus is retained.

It is known that the HN protein in the tunica externa of the Sendai virus recognizes the receptor (possessing sialic acid at the terminal of sugar chain) on the cell membrane and adsorbs it, leading to the induction of membrane fusion mediated by F protein (another component of the envelope). The genomic RNA of the Sendai virus contained in HVJ-E has been inactivated completely and has neither infective nor proliferative potentials in humans or experimental animals. HVJ-E can be used safely at ordinary laboratories, without requiring any special operations or facilities.
*HVJ:Hemagglutinating Virus of Japan.


Conventional non-viral transfection tools, including cationic lipids, are incorporated into cells through endocytosis which results in degradation of the transferred DNA by lysosomes.
On the other hand, HVJ Envelope VECTOR resists degradation by lysosomes, making it easy to transfer the specified DNA. Therefore, HVJ Envelope VECTOR yields highly efficient gene expression.
Sialic acid receptors, which are needed to trigger binding with HN protein, exist in almost all animal cells. Thus, HVJ Envelope VECTOR is useful for a wide range of targets.

Features and Advantages:

GenomONE-Neo
unique, exciting transfection kit for cells, tissue, and live animals. 

Use GenomONE-Neo: for: 

siRNA
strong knockdowns with low pM amounts of siRNA. Chemical modification of siRNA not required; means high functional expression..

DNA
small and large, linear or circular

Oligonucleotides
DNA, RNA

Proteins
enzymes and antibodies retain function and specificity

In vivo transfection!!
protocol and literature citations for effectiveness of molecules transfected into live animals following direct injection of HVJ-E vector.

High throughput screening
transfect multiple molecules simultaneously

1. Wide usability
GenomONE HVJ Envelope VECTOR KIT is a highly flexible tool for transfecting a wide variety of molecules (plasmid DNAs, siRNAs, oligonucleotides, proteins, antibodies etc.) into cultured cells (adherent and non-adherent) and tissue. GenomONE is useful for transfecting sensitive primary cells and is further distinguished by its ability to deliver contents into live animals. Many literature citations are available for each GenomONE-Neo application.
 

2. Safety
Unlike cationic lipids, GenomONE-Neo delivers the molecule of interest into cells via membrane fusion, not by endocytosis where cargo may be degraded by lysosomal enzymes. Since GenomeONE-Neo is a purified HVJ (Sendai virus) product, prepared from virus particles completely inactivated for infectious ability and proliferative potential it is completely safe to use without special precautions.
 

3. Easy
GenomONE provides ready-to-use kits containing the HVJ envelope and required auxiliary reagents (incorporation enhancer, incorporation reagent, introduction enhancer, and buffer). Suggested protocols for all major applications are included.

Comparison data of GenomONE-Neo with cationic lipid-based reagents (liposome):

  1. Comparison of GenomONE ™ -Neo with cationic lipid-based reagent for siRNA delivery
1. Phospholamban (PLB) gene knockdown in primary rat myocardial cell cultures
  GenomONE-Neo

 

1:Non-treated
2:PLB siRNA (10μg )
3:PLB siRNA (2μg)
4:Scramble siRNA (10μg)
5:HVJ-E only

Cationic lipid reagent

 

1:PLB siRNA (10μg )
2:PLB siRNA (2μg)
3:Scramble siRNA (10μg )
4:Non-treated

The efficiency of siRNA transfer into cells was high (80-100%) with both reagents.  However, compared to cationic lipid, GenomONE-Neo  induced knock-down of the target protein more efficiently at lower concentrations of siRNA.
 

【Data】Drs. M. Arai and A. Watanabe (Gunma University Graduate School of Medicine)
【Related article】Watanabe et al., J. Mol. Cell. Cardiol., 37 (3), 691-698 (2004)

   
  2 Bin1 gene knockdown in C2C12 (mouse myoblast cell line) after induction of differentiation
 
Use of conventional representative siRNA transfection reagents (products A, B, and C) failed to exert sufficient knock-down effects, demonstrating the superiority of GenomONE-Neo  to these products.
 

【Data】 Drs. C. Kojima* and H. Sabe (Osaka Bioscience Institute, *Current address: Osaka Prefecture University)
【Related article】Kojima et al.,  EMBO J., 23, 4413-4422 (2004).

   
  3 Eg5 gene knockdown in various cell cultures

 
Eg5-siRNA was incorporated in each reagent to a final concentration of 50 nM in a 96-well plate. Cells of various lines were treated with the reagents. Forty-eight hours later, the percentage of viable cells was measured by WST-1 assay. Eg5 knockdown resulted in suppression of cell proliferation and induced apoptosis. The finding of a lower percentage of viable cells indicated stronger Eg5 knockdown effects.
   
  4 EGFP gene knockdown in monkey ES cells
Biochem. Biophys. Res. Commun., 331, 1039-1044 (2005).
2. Highly efficient in vivo gene transfer to the mouse uterus
-Comparison data with cationic lipid-based reagent-
Mol. Hum. Reprod., 9, 603-609 (2003)

3.Transfection of neuroprogenitor cells with iron nanoparticles for magnetic resonance imaging tracking
-Comparison data with cationic lipid-based reagent-
Mol. Imaging Biol., 7, 286-295 (2005). 

Kit component:

GenomONE™-Neo EX Kit Contents GN001EX GN004EX
Freeze-dried HVJ-E 0.25 ml 1 tube 4 tubes
reagent A 0.5 ml 1 tube 1 tube
reagent B 0.3 ml 1 tube 1 tube
reagent C 1.0 ml 1 tube 1 tube
Buffer 6.5ml 1 tube 1 tube

The HVJ Envelope VECTOR KIT "GenomONE-Neo EX" series consists of HVJ Envelope and sub reagent set. Sub reagent set consists of 3 reagents (reagent A, reagent B and reagent C) and buffer for suspension or dilution.

  • Since the HVJ envelope of GenomONE-Neo EX (HVJ-E(N)) has a high fusing capability, it allows completion of transfection through a shorter contact duration with cells or tissues and has more suitable characteristics as follows: the reduction of toxicity to sensitive primary cultured cells, and the use in vivo where the transfected molecule is likely to diffuse due to inter-tissue transfer.
  • Some cultured cell lines show intense cell-to-cell fusion when GenomONE-Neo EX is used.
  • Applicability of  GenomONE-Neo EX may vary depending on the type of cell and the features of the experimental system. 

_____________________________________________________________________________________________________________________________

GenomONE-Neo EX HVJ-E 1 vial Transfection Reagents ISK-GN-001-EX 1 SET €262
GenomONE-Neo EX HVJ-E 4 vial(s) Transfection Reagents ISK-GN-004-EX 1 SET €600
GenomONE-Neo EX HVJ-E 40 vial(s) Transfection Reagents ISK-GN-040-EX 1 SET €3485

 

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