Non-integrated lentivirus - a new genetic research tool

2021-08-11 01:03:49
Non-integrated lentivirus - a new genetic research tool
Conventional lentiviral vectors can efficiently infect dividing cells and non-dividing cells, and effectively integrate foreign genes into the host chromosome to achieve persistent expression. However, this random integration poses the risk of insertional mutations. Therefore, researchers also expect to use a non-integrated lentiviral packaging system.

Bainwei Biosystems mutated the integrase protein and introduced an integrase-deficient lentiviral packaging system (non-integrated lentivirus). Non-integrating lentiviruses produce cyclic episomes in the cells of interest that are transiently expressed in dividing cells but stably expressed in non-dividing cells. Compared with the integrated lentivirus, the non-integrated lentivirus leads to a greatly reduced risk of insertional mutation, and is suitable for iPS research, RNA interference, homologous recombination and the like.

The non-integrated lentiviral packaging system belongs to the fourth generation of packaging system, which also separates the pol gene, making gag, pol and env three independent carriers instead of two carriers, thus greatly reducing the ability to produce replication. The possibility of viruses further enhances biosecurity. This system can be used with other lentiviral vectors to prepare high titer lentiviral particles that can reach 5x10 8 IU/ml without concentration. There are three main factors that can be used to prepare high titer virus particles.

First, the proportion of each vector in the non-integrated lentiviral system is optimized to greatly increase the amount of protein required for viral packaging. Secondly, the Tet-Off transcriptional activator (tTA) was introduced to form a cascade-enlarged expression effect of the viral packaging protein, which greatly increased the expression level of the viral packaging protein. Third, Bainwei Bio has transfection reagents with high transfection efficiency, which can efficiently transfect Lenti-X 293T packaging cell lines to prepare high titer lentiviral particles. This system is compatible with any lentiviral expression vector.

The main advantages of non-integrated lentivirus:
• Efficient gene transduction, does not insert into the host genome • Low risk of insertional mutations • Use of VSV-G envelope, broad host range • Transient expression in dividing cells, stable expression in non-dividing cells • With any lentiviral vector Used together to produce a non-integrated lentivirus • Produce a high titer lentivirus
In short, non-integrated lentiviruses are highly effective gene research tools because they express high expression genes without causing mutations.
As a leader in the field of viral packaging, Bai Enwei also has retroviruses, which are ideal for genetic research technology services.

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