Gene editing facility is established in July 2016, mainly composed of one molecular laboratory and one secondary biosafety laboratory (BSL-2). The work of our facility is to provide services for the research of non-human primate disease model by package and purification of viral gene delivery vectors.
Viral gene delivery vectors are important tools in scientific research. They are widely used because the low cost and long-term stable gene expression. The most commonly used systems are lentivirus and adeno-associated virus (AAV).
1) The lentivirus vector was engineered from human immunodeficiency virus (HIV), it can infect both mitotic and non-mitotic cells. Through lentivirus infection, target genes can be integrated into host chromosome and express constantly. Lentivirus can effectively infect neuron cells, liver cells, myocardial cells, tumor cells, endothelial cells, stem cells and other types of cells, etc. Compared with other viral vectors, lentivirus has larger gene capacity. The largest DNA fragment loading is 8-9 KB, while adeno-associated virus is only 4 KB.
2) Adeno associated virus (AAV) is a replication deficient virus whose replication requires the assistance of adenovirus or herpesvirus. At present, AAV helper free system can produce recombinant human adeno-associated virus without helper virus. AAV can infect both mitotic and non-mitotic cells. In humans and mice, AAV infection can mediate the directional integration of foreign genes at low frequency (no more than 2%). The virus genome forms an accompaniment in the infected cells without integration, which can replicate with chromosomes. Therefore, both the integrated exogenous DNA and the non-integrated exogenous DNA can replicate and transcribe stably in the nucleus, producing RNA products and protein products. It can effectively infect a variety of tissue types, and its immunogenicity is very low. At present, there are no reports of human and mammalian diseases caused by AAV. Especially in the overall level of research, compared with other commonly used virus tools, AAV virus has small particles, good diffusion performance, and long-term expression ability (> 6 months).
