Generation of a self‐cleaved inducible Cre recombinase for efficient temporal genetic manipulation
- 14 January 2020
- journal article
- research article
- Published by Springer Science and Business Media LLC in The EMBO Journal
- Vol. 39 (4), e102675
- https://doi.org/10.15252/embj.2019102675
Abstract
Site‐specific recombinase‐mediated genetic technology, such as inducible Cre‐loxP recombination (CreER), is widely used for in vivo genetic manipulation with temporal control. The Cre‐loxP technology improves our understanding on the in vivo function of specific genes in organ development, tissue regeneration, and disease progression. However, inducible CreER often remains inefficient in gene deletion. In order to improve the efficiency of gene manipulation, we generated a self‐cleaved inducible CreER (sCreER) that switches inducible CreER into a constitutively active Cre by itself. We generated endocardial driver Npr3‐sCreER and fibroblast driver Col1a2‐sCreER, and compared them with conventional Npr3‐CreER and Col1a2‐CreER, respectively. For easy‐to‐recombine alleles such as R26‐tdTomato, there was no significant difference in recombination efficiency between sCreER and the conventional CreER. However, for alleles that were relatively inert for recombination such as R26‐Confetti, R26‐LZLT, R26‐GFP, or VEGFR2flox/flox alleles, sCreER showed a significantly higher efficiency in recombination compared with conventional CreER in endocardial cells or fibroblasts. Compared with conventional CreER, sCreER significantly enhances the efficiency of recombination to induce gene expression or gene deletion, allowing temporal yet effective in vivo genomic modification for studying gene function in specific cell lineages.Keywords
Funding Information
- China Association for Science and Technology (2018QNRC001, 2017QNRC001)
- Boehringer Ingelheim
- National Natural Science Foundation of China (31730112, 91639302, 31625019, 91849202, 81761138040, 81872241, 31701292, 31801215, 31922032)
- China Postdoctoral Science Foundation
- AstraZeneca
This publication has 38 references indexed in Scilit:
- Subepicardial endothelial cells invade the embryonic ventricle wall to form coronary arteriesCell Research, 2013
- Lost in TransgenesisCirculation Research, 2012
- Tamoxifen Induces Rapid, Reversible Atrophy, and Metaplasia in Mouse StomachGastroenterology, 2012
- Mosaic Analysis with Double Markers Reveals Tumor Cell of Origin in GliomaCell, 2011
- Intestinal Crypt Homeostasis Results from Neutral Competition between Symmetrically Dividing Lgr5 Stem CellsCell, 2010
- A robust and high-throughput Cre reporting and characterization system for the whole mouse brainNature Neuroscience, 2009
- Alternatively spliced vascular endothelial growth factor receptor-2 is an essential endogenous inhibitor of lymphatic vessel growthNature Medicine, 2009
- Reassessment of Isl1 and Nkx2-5 cardiac fate maps using a Gata4-based reporter of Cre activityDevelopmental Biology, 2008
- Tamoxifen exposure and risk of oesophageal and gastric adenocarcinoma: a population-based cohort study of breast cancer patients in SwedenBritish Journal of Cancer, 2006
- Regulation of Cre Recombinase Activity by Mutated Estrogen Receptor Ligand-Binding DomainsBiochemical and Biophysical Research Communications, 1997