Conditional transgenesis using Dimerizable Cre (DiCre).
Cre recombinase is extensively used to engineer the genome of experimental animals. However, its usefulness is still limited by the lack of an efficient temporal control over its activity. We have recently developed a conceptually new approach to regulate Cre recombinase, that we have called Dimeriz...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2007-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC2131782?pdf=render |
| _version_ | 1819099045954584576 |
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| author | Nicolas Jullien Isabelle Goddard Samia Selmi-Ruby Jean-Luc Fina Harold Cremer Jean-Paul Herman |
| author_facet | Nicolas Jullien Isabelle Goddard Samia Selmi-Ruby Jean-Luc Fina Harold Cremer Jean-Paul Herman |
| author_sort | Nicolas Jullien |
| collection | DOAJ |
| description | Cre recombinase is extensively used to engineer the genome of experimental animals. However, its usefulness is still limited by the lack of an efficient temporal control over its activity. We have recently developed a conceptually new approach to regulate Cre recombinase, that we have called Dimerizable Cre or DiCre. It is based on splitting Cre into two inactive moieties and fusing them to FKBP12 (FK506-binding protein) and FRB (binding domain of the FKBP12-rapamycin associated protein), respectively. These latter can be efficiently hetero-dimerized by rapamycin, leading to the reinstatement of Cre activity. We have been able to show, using in vitro approaches, that this ligand-induced dimerization is an efficient way to regulate Cre activity, and presents a low background activity together with a high efficiency of recombination following dimerization. To test the in vivo performance of this system, we have, in the present work, knocked-in DiCre into the Rosa26 locus of mice. To evaluate the performance of the DiCre system, mice have been mated with indicator mice (Z/EG or R26R) and Cre-induced recombination was examined following activation of DiCre by rapamycin during embryonic development or after birth of progenies. No recombination could be observed in the absence of treatment of the animals, indicating a lack of background activity of DiCre in the absence of rapamycin. Postnatal rapamycin treatment (one to five daily injection, 10 mg/kg i.p) induced recombination in a number of different tissues of progenies such as liver, heart, kidney, muscle, etc. On the other hand, recombination was at a very low level following in utero treatment of DiCrexR26R mice. In conclusion, DiCre has indeed the potentiality to be used to establish conditional Cre-deleter mice. An added advantage of this system is that, contrary to other modulatable Cre systems, it offers the possibility of obtaining regulated recombination in a combinatorial manner, i.e. induce recombination at any desired time-point specifically in cells characterized by the simultaneous expression of two different promoters. |
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| id | doaj.art-406d82ef7df74fc7b482ed34f62aebf1 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-22T00:40:39Z |
| publishDate | 2007-01-01 |
| publisher | Public Library of Science (PLoS) |
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| series | PLoS ONE |
| spelling | doaj.art-406d82ef7df74fc7b482ed34f62aebf12022-12-21T18:44:42ZengPublic Library of Science (PLoS)PLoS ONE1932-62032007-01-01212e135510.1371/journal.pone.0001355Conditional transgenesis using Dimerizable Cre (DiCre).Nicolas JullienIsabelle GoddardSamia Selmi-RubyJean-Luc FinaHarold CremerJean-Paul HermanCre recombinase is extensively used to engineer the genome of experimental animals. However, its usefulness is still limited by the lack of an efficient temporal control over its activity. We have recently developed a conceptually new approach to regulate Cre recombinase, that we have called Dimerizable Cre or DiCre. It is based on splitting Cre into two inactive moieties and fusing them to FKBP12 (FK506-binding protein) and FRB (binding domain of the FKBP12-rapamycin associated protein), respectively. These latter can be efficiently hetero-dimerized by rapamycin, leading to the reinstatement of Cre activity. We have been able to show, using in vitro approaches, that this ligand-induced dimerization is an efficient way to regulate Cre activity, and presents a low background activity together with a high efficiency of recombination following dimerization. To test the in vivo performance of this system, we have, in the present work, knocked-in DiCre into the Rosa26 locus of mice. To evaluate the performance of the DiCre system, mice have been mated with indicator mice (Z/EG or R26R) and Cre-induced recombination was examined following activation of DiCre by rapamycin during embryonic development or after birth of progenies. No recombination could be observed in the absence of treatment of the animals, indicating a lack of background activity of DiCre in the absence of rapamycin. Postnatal rapamycin treatment (one to five daily injection, 10 mg/kg i.p) induced recombination in a number of different tissues of progenies such as liver, heart, kidney, muscle, etc. On the other hand, recombination was at a very low level following in utero treatment of DiCrexR26R mice. In conclusion, DiCre has indeed the potentiality to be used to establish conditional Cre-deleter mice. An added advantage of this system is that, contrary to other modulatable Cre systems, it offers the possibility of obtaining regulated recombination in a combinatorial manner, i.e. induce recombination at any desired time-point specifically in cells characterized by the simultaneous expression of two different promoters.http://europepmc.org/articles/PMC2131782?pdf=render |
| spellingShingle | Nicolas Jullien Isabelle Goddard Samia Selmi-Ruby Jean-Luc Fina Harold Cremer Jean-Paul Herman Conditional transgenesis using Dimerizable Cre (DiCre). PLoS ONE |
| title | Conditional transgenesis using Dimerizable Cre (DiCre). |
| title_full | Conditional transgenesis using Dimerizable Cre (DiCre). |
| title_fullStr | Conditional transgenesis using Dimerizable Cre (DiCre). |
| title_full_unstemmed | Conditional transgenesis using Dimerizable Cre (DiCre). |
| title_short | Conditional transgenesis using Dimerizable Cre (DiCre). |
| title_sort | conditional transgenesis using dimerizable cre dicre |
| url | http://europepmc.org/articles/PMC2131782?pdf=render |
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