REGULATORY ELEMENTS IN THE INTRONS OF THE HUMAN HPRT GENE ARE NECESSARY FOR ITS EXPRESSION IN EMBRYONIC STEM-CELLS

被引：90

作者：

REID, LH ^{[1
]}

GREGG, RG ^{[1
]}

SMITHIES, O ^{[1
]}

KOLLER, BH ^{[1
]}

机构：

[1] UNIV WISCONSIN,DEPT MED GENET,MADISON,WI 53706

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 1990年 / 87卷 / 11期

关键词：

Enhancer; Gene expression; Gene targeting;

D O I：

10.1073/pnas.87.11.4299

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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 somotic cell lines failed to confer hypoxanthine/aminopterin/ thymicine (HAT) resistance in ES cells. In contrast, constructs containing introns l 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.

引用

页码：4299 / 4303

页数：5

共 31 条

[1] EVIDENCE FOR A COMPLEX REGULATORY ARRAY IN THE 1ST INTRON OF THE HUMAN ADENOSINE-DEAMINASE GENE [J].