Cre reconstitution allows for DNA recombination selectively in dual-marker-expressing cells in transgenic mice.

@article{citeulike:1749973, abstract = {Cre/LoxP-based DNA recombination has been used to introduce desired DNA rearrangements in various organisms, having for example, greatly assisted genetic analyses in mice. For most applications, single gene promoters are used to drive Cre production for conditional gene activation/inactivation or lineage-tracing experiments. Such a manipulation introduces Cre in all cells in which the utilized promoter is active. To overcome the limited selectivity of single promoters for cell-type-specific recombination, we have explored the 'dual promoter combinatorial control' of Cre activity, so that Cre activity could be restricted to cells that express dual protein markers. We efficiently reconstituted Cre activity from two modified, inactive Cre fragments. Cre re-association was greatly enhanced by fusing the Cre fragments separately to peptides that can form a tight antiparallel leucine zipper. The co-expressed Cre fusion fragments showed substantial activity in cultured cells. As proof of principle of the utility of this technique in vivo for manipulating genes specifically in dual-marker-positive cells, we expressed each inactive Cre fragments in transgenic mice via individual promoters. Result showed the effective reconstitution of Cre activates LoxP recombination in the co-expressing cells.}, address = {Program in Developmental Biology and Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, TN 37232, USA, Department of Medicine and Therapeutics, Prince of Wales Hospital, The Chinese University of Hong Kong, Shatin, Hong Kong and Center for AIDS Health Disparities Research, Meharry Medical College, Nashville, TN 37208, USA.}, author = {Xu, Yanwen and Xu, Gang and Liu, Bindong and Gu, Guoqiang }, citeulike-article-id = {1749973}, issn = {1362-4962}, journal = {Nucleic Acids Res}, keywords = {cre, dual, flox, lineage, marker, monocyte, promoter, recombination, system, tracing, track, transgenic}, month = {September}, posted-at = {2008-08-08 18:28:10}, priority = {2}, title = {Cre reconstitution allows for DNA recombination selectively in dual-marker-expressing cells in transgenic mice.}, url = {http://view.ncbi.nlm.nih.gov/pubmed/17893102}, year = {2007} }

TY - JOUR ID - citeulike:1749973 L3 - citeulike-article-id:1749973 TI - Cre reconstitution allows for DNA recombination selectively in dual-marker-expressing cells in transgenic mice. JF - Nucleic Acids Res AD - Program in Developmental Biology and Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, TN 37232, USA, Department of Medicine and Therapeutics, Prince of Wales Hospital, The Chinese University of Hong Kong, Shatin, Hong Kong and Center for AIDS Health Disparities Research, Meharry Medical College, Nashville, TN 37208, USA. SN - 1362-4962 N2 - Cre/LoxP-based DNA recombination has been used to introduce desired DNA rearrangements in various organisms, having for example, greatly assisted genetic analyses in mice. For most applications, single gene promoters are used to drive Cre production for conditional gene activation/inactivation or lineage-tracing experiments. Such a manipulation introduces Cre in all cells in which the utilized promoter is active. To overcome the limited selectivity of single promoters for cell-type-specific recombination, we have explored the 'dual promoter combinatorial control' of Cre activity, so that Cre activity could be restricted to cells that express dual protein markers. We efficiently reconstituted Cre activity from two modified, inactive Cre fragments. Cre re-association was greatly enhanced by fusing the Cre fragments separately to peptides that can form a tight antiparallel leucine zipper. The co-expressed Cre fusion fragments showed substantial activity in cultured cells. As proof of principle of the utility of this technique in vivo for manipulating genes specifically in dual-marker-positive cells, we expressed each inactive Cre fragments in transgenic mice via individual promoters. Result showed the effective reconstitution of Cre activates LoxP recombination in the co-expressing cells. KW - cre KW - dual KW - flox KW - lineage KW - marker KW - monocyte KW - promoter KW - recombination KW - system KW - tracing KW - track KW - transgenic AU - Xu, Yanwen AU - Xu, Gang AU - Liu, Bindong AU - Gu, Guoqiang PY - 2007/09/24/ UR - http://view.ncbi.nlm.nih.gov/pubmed/17893102 ER -