Regulatory elements in the introns of the human HPRT gene are necessary for its expression in embryonic stem cells.
AUTOR(ES)
Reid, L H
RESUMO
We have examined the expression of transfected human hypoxanthine phosphoribosyltransferase minigenes (HPRT) in mouse embryonic stem (ES) cells. cDNA constructs of this gene that have been successfully used in somatic cell lines failed to confer hypoxanthine/aminopterin/thymidine (HAT) resistance in ES cells. In contrast, constructs containing introns 1 and 2 from the HPRT gene produced a high frequency of HAT-resistant colonies. This observation allowed us to identify two sequences in these introns that influence expression of the HPRT gene in ES cells. One element, located in intron 2, is required for effective HPRT expression in these cells; the other element, located in intron 1, acts as an enhancer of HPRT expression. Using this information, we have constructed an HPRT minigene that can be used for either positive or negative selection in ES cell experiments. This dual capability allows the design of "in-out" procedures to create subtle changes in target genes by homologous recombination with the aid of this selectable minigene.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=54096Documentos Relacionados
- Targeted mutation of the Hprt gene in mouse embryonic stem cells.
- The length of homology required for gene targeting in embryonic stem cells.
- Oxygen supply and oxygen-dependent gene expression in differentiating embryonic stem cells.
- Embryonic stem cell virus, a recombinant murine retrovirus with expression in embryonic stem cells.
- A lacZ-hygromycin fusion gene and its use in a gene trap vector for marking embryonic stem cells.
