Efficient deletion of LoxP-flanked selectable marker genes from the genome of transgenic pigs by an engineered Cre recombinase.
Transgenic Res
; 29(3): 307-319, 2020 06.
Article
en En
| MEDLINE
| ID: mdl-32410183
ABSTRACT
Genetically modified (GM) pigs hold great promises for pig genetic improvement, human health and life science. When GM pigs are produced, selectable marker genes (SMGs) are usually introduced into their genomes for host cell or animal recognition. However, the SMGs that remain in GM pigs might have multiple side effects. To avoid the possible side effects caused by the SMGs, they should be removed from the genome of GM pigs before their commercialization. The Cre recombinase is commonly used to delete the LoxP sites-flanked SMGs from the genome of GM animals. Although SMG-free GM pigs have been generated by Cre-mediated recombination, more efficient and cost-effective approaches are essential for the commercialization of SMG-free GM pigs. In this article we describe the production of a recombinant Cre protein containing a cell-penetrating and a nuclear localization signal peptide in one construct. This engineered Cre enzyme can efficiently excise the LoxP-flanked SMGs in cultured fibroblasts isolated from a transgenic pig, which then can be used as nuclear donor cells to generate live SMG-free GM pigs harboring a desired transgene by somatic cell nuclear transfer. This study describes an efficient and far-less costly method for production of SMG-free GM pigs.
Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Recombinación Genética
/
Ingeniería Genética
/
Marcadores Genéticos
/
Genoma
/
Transgenes
/
Integrasas
/
Técnicas de Transferencia Nuclear
Límite:
Animals
Idioma:
En
Año:
2020
Tipo del documento:
Article