{Reference Type}: Journal Article
{Title}: Platform establishment of the Cre-loxP recombination system for genetic manipulation of the Lumpy skin disease virus.
{Author}: Ren S;Zhang Y;Gao X;Wang X;Tong L;Wang S;Sun Y;Yin X;Chen H;
{Journal}: Vet Microbiol
{Volume}: 294
{Issue}: 0
{Year}: 2024 Jul 18
{Factor}: 3.246
{DOI}: 10.1016/j.vetmic.2024.110122
{Abstract}: Lumpy skin disease virus (LSDV) is a rapidly emerging pathogen in Asia, including China. Genetic manipulation of the LSDV is essential for the elucidation of the pathogenic mechanism and biological function of the LSDV-encoded protein. In this study, we established a platform for the Cre-loxP recombination system under a modified early-late H5 promoter of the VACV for quick construction of the recombinant LSDV virus. The recombinant virus, LSDV-EGFP-ΔTK, was purified and obtained using serial limited dilution and picking the single cells methods. Using the lentiviral package system, a Cre recombinase enzyme stable expression MDBK cell line was established to supply the Cre recombinase for the reporter gene excision. A genetically stable, safe TK gene-deleted LSDV (LSDV-ΔTK) was constructed using homologous recombination and the Cre-loxP system. It was purified using limited dilution in the MDBK-Cre cell line. Establishing the Cre-loxP recombination system will enable sequential deletion of the interested genes from the LSDV genome and genetic manipulation of the LSDV genome, providing technical support and a platform for developing the attenuated LSDV vaccine.