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PMID: 17209010 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Controlled expression of transgenes introduced by in vivo electroporation.

Matsuda T, Cepko CL

Abstract

In vivo electroporation is a powerful technique for the introduction of genes into organisms. Temporal and spatial regulation of expression of introduced genes, or of RNAi, would further enhance the utility of this method. Here we demonstrate conditional regulation of gene expression from electroporated plasmids in the postnatal rat retina and the embryonic mouse brain. For temporal regulation, Cre/loxP-mediated inducible expression vectors were used in combination with a vector expressing a conditionally active form of Cre recombinase, which is activated by 4-hydroxytamoxifen. Onset of gene expression was regulated by the timing of 4-hydroxytamoxifen administration. For spatial regulation, transgenes were expressed by using promoters specific for rod photoreceptors, bipolar cells, amacrine cells, Müller glia or progenitor cells. Combinations of these constructs will facilitate a variety of experiments, including cell-type-specific gene misexpression, conditional RNAi, and fate mapping of progenitor and precursor cells.

MeSH Terms
Animals Basic Helix-Loop-Helix Transcription Factors/genetics,metabolism Cell Line Cell Lineage Electroporation/methods Gene Expression/genetics Gene Expression Regulation Homeodomain Proteins/genetics,metabolism Humans Integrases/genetics,metabolism Promoter Regions, Genetic/genetics RNA Interference Rats Retina/cytology,metabolism Transcription Factor HES-1 Transfection/methods
Chemicals
Basic Helix-Loop-Helix Transcription Factors Hes1 protein, rat Homeodomain Proteins RAX protein, rat Transcription Factor HES-1 Cre recombinase Integrases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Matsuda Takahiko
Department of Genetics and Howard Hughes Medical Institute, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, MA 02115, USA.
Cepko Constance L
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
2007-01-16
Epub
2007-00-05
Pages
1027-32
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC1764220
Subset
IM
Grants
NEI NIH HHS · EY 08064 · United States
NEI NIH HHS · EY 09676 · United States
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