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In the biomedical field, the transfer of plasmid vectors into mammalian cells using conventional transfection is the most widely used method. Although a number of more advanced gene introduction vector systems have been developed in recent years (e.g., lentiviral vectors, adenoviral vectors, AAV vectors, and piggyBac, etc.), conventional plasmid gene expression vectors are still used in the majority of laboratories, mainly due to their technical simplicity and efficient transfection rates in various types of cells.

Lentivirus vector is a gene therapy vector developed on the base of HIV-1 (human immunodeficiency virus type 1). Lentivirus vector is a powerful tool for introduction of exogenous genes, with the advantages of wide infection spectrum, effective infection and long-term stable expression of exogenous genes. Now the lentivirus system has been widely used in various cell lines for gene overexpression, RNA interference, microRNA research, as well as in vivo experiments.

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Adeno-Associated Virus (AAV package) belongs to the family of parvovirus and is a non-enveloped single-stranded DNA virus, with a genome size of 4.7 kb composed of two open reading frames (ORFs) flanked by inverted terminal repeat sequences (ITRs) of 145 nucleotides each. AAV vectors are one of the most commonly used vectors for a variety of gene therapy applications and have been widely used in clinics and basic research mainly owing to their capable of transducing a wide range of species and tissues in vivo with no evidence of toxicity, and it generates relatively mild innate and adaptive immune responses as well as their long-term transgene expression and steady physical properties.

Seamless cloning technique from HANBIO is a new, fast and simple cloning method, which can insert one or more target DNA fragments at any site of the plasmid without any restriction enzymes and ligases. Breaking through the traditional double enzyme digestion and linking, the recombinant vector of high efficiency cloning can be obtained by one-step recombinant method, which greatly improves the working efficiency.

To create a lentivirus stable cell line, cells are typically transduced with lentiviral particles containing the desired gene of interest. The lentiviral vector integrates into the host cell's genome, leading to stable expression of the gene in the cell line. This allows researchers to study the effects of the gene of interest in a controlled and reproducible manner.