A regulatory toolbox of MiniPromoters to drive selective expression in the brain

doi:10.1073/pnas.1009158107

A regulatory toolbox of MiniPromoters to drive selective expression in the brain

^aCentre for Molecular Medicine and Therapeutics at the Child and Family Research Institute, University of British Columbia, Vancouver, BC, Canada V5Z4H4;
^bDepartment of Anatomy and Neurobiology, University of Tennessee, Memphis, TN 38163;
^cDepartment of Medical Genetics, University of British Columbia, Vancouver, BC, Canada V5Z4H4;
^dGenetics Graduate Program, University of British Columbia, Vancouver, BC, Canada V5Z4H4;
^eCanada's Michael Smith Genome Sciences Centre, British Columbia Cancer Agency, Vancouver, BC, Canada V5Z4S6;
^fDepartment of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, BC, Canada V5A1S6; and
^gDepartment of Psychiatry, University of British Columbia, Vancouver, BC, Canada V6T2A1

Communicated by Richard L. Sidman, Beth Israel Deaconess Medical Center, Boston, MA, June 28, 2010 (received for review January 15, 2010)
↵¹D.J.S. and L.L. contributed equally to this work.
↵²C.N.d.L. and K.G.B. contributed equally to this work.

Abstract

The Pleiades Promoter Project integrates genomewide bioinformatics with large-scale knockin mouse production and histological examination of expression patterns to develop MiniPromoters and related tools designed to study and treat the brain by directed gene expression. Genes with brain expression patterns of interest are subjected to bioinformatic analysis to delineate candidate regulatory regions, which are then incorporated into a panel of compact human MiniPromoters to drive expression to brain regions and cell types of interest. Using single-copy, homologous-recombination “knockins” in embryonic stem cells, each MiniPromoter reporter is integrated immediately 5′ of the Hprt locus in the mouse genome. MiniPromoter expression profiles are characterized in differentiation assays of the transgenic cells or in mouse brains following transgenic mouse production. Histological examination of adult brains, eyes, and spinal cords for reporter gene activity is coupled to costaining with cell-type–specific markers to define expression. The publicly available Pleiades MiniPromoter Project is a key resource to facilitate research on brain development and therapies.

Footnotes

³To whom correspondence should be addressed. E-mail: simpson{at}cmmt.ubc.ca.
Author contributions: S.J.M.J., R.A.H., D.G., W.W.W., and E.M.S. designed research; E.P.-C., D.J.S., L.L., C.N.d.L., K.G.B., S.J.H.-S., D.L.F., J.A., M.A., D.J.A., N.B., S.F.B., R.J.B., E.B., T.R.C., M.C., J.C., Y.C., J.C.Y.C., V.C., T.R.D., L.D., C.A.D., E.K.F., R.G., K.H., T.G.H., B.I., S.J., S.K.-z., I.K., S. Laprise, N.Y.L., J.S.L, S. Lithwick, F.L., J.L, M.L., M. McConechy, A.J.M., M. Milisavljevic, J.M., K.O., B.P., D.L.P., J.-F.S., M.I.S., B.T., A.T., J.L.T., R.V., J.V., R.F.W., G.W., B.K.Y.W., S.H.W., T.Y.T.W., G.S.Y., and A.R.Y. performed research; E.P.-C., D.J.S., L.L., C.N.d.L., K.G.B., S.J.H.-S., D.L.F., S.J.M.J., R.A.H., D.G., W.W.W., and E.M.S. analyzed data; and E.P.-C. wrote the paper.
The authors declare no conflict of interest.
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