Altering the expression in mice of genes by modifying their 3' regions.
Polymorphic differences altering expression of genes without changing their products probably underlie human quantitative traits affecting risks of serious diseases, but methods for investigating such quantitative differences in animals are limited. Accordingly, we have developed a procedure for changing the expression in mice of chosen genes over a 100-fold range while retaining their chromosomal location and transcriptional controls. To develop the procedure, we first dissected the effects in embryonic stem (ES) cells of elements within and downstream of the 3' untranslated region (UTR) of a single copy transgene at the Hprt locus. As expected, protein expression varied with the steady-state level and half-life of the mRNA. The rank order of expression with various tested 3' regions is the same in ES cells, and in cardiomyocytes and trophoblastocytes derived from them. In mice having two functionally different native genes with modified 3'UTRs, the desired expression was obtained.
Kakoki
Masao
M
Department of Pathology and Laboratory Medicine, University of North Carolina, Chapel Hill, NC 27599, USA.
Tsai
Yau-Sheng
YS
Kim
Hyung-Suk
HS
Hatada
Seigo
S
Ciavatta
Dominic J
DJ
Takahashi
Nobuyuki
N
Arnold
Larry W
LW
Maeda
Nobuyo
N
Smithies
Oliver
O
D020413:3' Untranslated Regions / Q000235:genetics*; D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D005786:Gene Expression Regulation / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D023421:Models, Animal; D008967:Molecular Biology / Q000379:methods*; D016296:Mutagenesis / Q000235:genetics*; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism; D011110:Polymorphism, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism; D019076:Transgenes / Q000235:genetics; D014327:Trophoblasts / Q000166:cytology / Q000378:metabolism
D020413:3' Untranslated Regions; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins