A doxycycline- and light-inducible Cre recombinase mouse model for optogenetic genome editing

The experimental need to engineer the genome both in time and space, has led to the development of several photoactivatable Cre recombinase systems. However, the combination of inefficient and non-intentional background recombination has prevented thus far the wide application of these systems in bi...

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Veröffentlicht in:	Nature communications Jg. 13; H. 1; S. 6442 - 15
Hauptverfasser:	Vizoso, Miguel, E. J. Pritchard, Colin, Bombardelli, Lorenzo, van den Broek, Bram, Krimpenfort, Paul, Beijersbergen, Roderick L., Jalink, Kees, van Rheenen, Jacco
Format:	Journal Article
Sprache:	Englisch
Veröffentlicht:	London Nature Publishing Group UK 28.10.2022 Nature Publishing Group Nature Portfolio
Schlagworte:	13/1 13/106 13/107 13/109 13/31 13/44 13/51 14 14/1 14/19 14/69 38 59 631/1647/2253 631/1647/334/1874/345 631/61/17/1511 631/67/2321 64 64/60 Animals Antibiotics Biomedical engineering Biomedical materials Biomedical research Cre recombinase Dermis Doxycycline Doxycycline - pharmacology Editing Engineers Gene Editing Gene expression Genome editing Genomes Humanities and Social Sciences Integrases - genetics Integrases - metabolism Medical research Mice Mice, Inbred Strains Mice, Transgenic multidisciplinary Mutagenesis Optogenetics Organoids Recombination Science Science (multidisciplinary) Tracing
ISSN:	2041-1723, 2041-1723
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Abstract	The experimental need to engineer the genome both in time and space, has led to the development of several photoactivatable Cre recombinase systems. However, the combination of inefficient and non-intentional background recombination has prevented thus far the wide application of these systems in biological and biomedical research. Here, we engineer an optimized photoactivatable Cre recombinase system that we refer to as doxycycline- and light-inducible Cre recombinase (DiLiCre). Following extensive characterization in cancer cell and organoid systems, we generate a DiLiCre mouse line, and illustrated the biological applicability of DiLiCre for light-induced mutagenesis in vivo and positional cell-tracing by intravital microscopy. These experiments illustrate how newly formed HrasV12 mutant cells follow an unnatural movement towards the interfollicular dermis. Together, we develop an efficient photoactivatable Cre recombinase mouse model and illustrate how this model is a powerful genome-editing tool for biological and biomedical research. Achieving spatial control of gene expression is important. Here the authors report an optimised photoactivatable Cre recombinase system, doxycycline- and light-inducible Cre recombinase (DiLiCre), and generate a DiLiCre mouse line which they use for mutagenesis in vivo and positional cell-tracing.
AbstractList	The experimental need to engineer the genome both in time and space, has led to the development of several photoactivatable Cre recombinase systems. However, the combination of inefficient and non-intentional background recombination has prevented thus far the wide application of these systems in biological and biomedical research. Here, we engineer an optimized photoactivatable Cre recombinase system that we refer to as doxycycline- and light-inducible Cre recombinase (DiLiCre). Following extensive characterization in cancer cell and organoid systems, we generate a DiLiCre mouse line, and illustrated the biological applicability of DiLiCre for light-induced mutagenesis in vivo and positional cell-tracing by intravital microscopy. These experiments illustrate how newly formed HrasV12 mutant cells follow an unnatural movement towards the interfollicular dermis. Together, we develop an efficient photoactivatable Cre recombinase mouse model and illustrate how this model is a powerful genome-editing tool for biological and biomedical research. Achieving spatial control of gene expression is important. Here the authors report an optimised photoactivatable Cre recombinase system, doxycycline- and light-inducible Cre recombinase (DiLiCre), and generate a DiLiCre mouse line which they use for mutagenesis in vivo and positional cell-tracing. The experimental need to engineer the genome both in time and space, has led to the development of several photoactivatable Cre recombinase systems. However, the combination of inefficient and non-intentional background recombination has prevented thus far the wide application of these systems in biological and biomedical research. Here, we engineer an optimized photoactivatable Cre recombinase system that we refer to as doxycycline- and light-inducible Cre recombinase (DiLiCre). Following extensive characterization in cancer cell and organoid systems, we generate a DiLiCre mouse line, and illustrated the biological applicability of DiLiCre for light-induced mutagenesis in vivo and positional cell-tracing by intravital microscopy. These experiments illustrate how newly formed HrasV12 mutant cells follow an unnatural movement towards the interfollicular dermis. Together, we develop an efficient photoactivatable Cre recombinase mouse model and illustrate how this model is a powerful genome-editing tool for biological and biomedical research.The experimental need to engineer the genome both in time and space, has led to the development of several photoactivatable Cre recombinase systems. However, the combination of inefficient and non-intentional background recombination has prevented thus far the wide application of these systems in biological and biomedical research. Here, we engineer an optimized photoactivatable Cre recombinase system that we refer to as doxycycline- and light-inducible Cre recombinase (DiLiCre). Following extensive characterization in cancer cell and organoid systems, we generate a DiLiCre mouse line, and illustrated the biological applicability of DiLiCre for light-induced mutagenesis in vivo and positional cell-tracing by intravital microscopy. These experiments illustrate how newly formed HrasV12 mutant cells follow an unnatural movement towards the interfollicular dermis. Together, we develop an efficient photoactivatable Cre recombinase mouse model and illustrate how this model is a powerful genome-editing tool for biological and biomedical research. The experimental need to engineer the genome both in time and space, has led to the development of several photoactivatable Cre recombinase systems. However, the combination of inefficient and non-intentional background recombination has prevented thus far the wide application of these systems in biological and biomedical research. Here, we engineer an optimized photoactivatable Cre recombinase system that we refer to as doxycycline- and light-inducible Cre recombinase (DiLiCre). Following extensive characterization in cancer cell and organoid systems, we generate a DiLiCre mouse line, and illustrated the biological applicability of DiLiCre for light-induced mutagenesis in vivo and positional cell-tracing by intravital microscopy. These experiments illustrate how newly formed HrasV12 mutant cells follow an unnatural movement towards the interfollicular dermis. Together, we develop an efficient photoactivatable Cre recombinase mouse model and illustrate how this model is a powerful genome-editing tool for biological and biomedical research. Achieving spatial control of gene expression is important. Here the authors report an optimised photoactivatable Cre recombinase system, doxycycline- and light-inducible Cre recombinase (DiLiCre), and generate a DiLiCre mouse line which they use for mutagenesis in vivo and positional cell-tracing. The experimental need to engineer the genome both in time and space, has led to the development of several photoactivatable Cre recombinase systems. However, the combination of inefficient and non-intentional background recombination has prevented thus far the wide application of these systems in biological and biomedical research. Here, we engineer an optimized photoactivatable Cre recombinase system that we refer to as doxycycline- and light-inducible Cre recombinase (DiLiCre). Following extensive characterization in cancer cell and organoid systems, we generate a DiLiCre mouse line, and illustrated the biological applicability of DiLiCre for light-induced mutagenesis in vivo and positional cell-tracing by intravital microscopy. These experiments illustrate how newly formed HrasV12 mutant cells follow an unnatural movement towards the interfollicular dermis. Together, we develop an efficient photoactivatable Cre recombinase mouse model and illustrate how this model is a powerful genome-editing tool for biological and biomedical research. Achieving spatial control of gene expression is important. Here the authors report an optimised photoactivatable Cre recombinase system, doxycycline- and light-inducible Cre recombinase (DiLiCre), and generate a DiLiCre mouse line which they use for mutagenesis in vivo and positional cell-tracing. The experimental need to engineer the genome both in time and space, has led to the development of several photoactivatable Cre recombinase systems. However, the combination of inefficient and non-intentional background recombination has prevented thus far the wide application of these systems in biological and biomedical research. Here, we engineer an optimized photoactivatable Cre recombinase system that we refer to as doxycycline- and light-inducible Cre recombinase (DiLiCre). Following extensive characterization in cancer cell and organoid systems, we generate a DiLiCre mouse line, and illustrated the biological applicability of DiLiCre for light-induced mutagenesis in vivo and positional cell-tracing by intravital microscopy. These experiments illustrate how newly formed HrasV12 mutant cells follow an unnatural movement towards the interfollicular dermis. Together, we develop an efficient photoactivatable Cre recombinase mouse model and illustrate how this model is a powerful genome-editing tool for biological and biomedical research.
ArticleNumber	6442
Author	E. J. Pritchard, Colin Beijersbergen, Roderick L. van den Broek, Bram van Rheenen, Jacco Jalink, Kees Vizoso, Miguel Bombardelli, Lorenzo Krimpenfort, Paul
Author_xml	– sequence: 1 givenname: Miguel orcidid: 0000-0002-9992-2851 surname: Vizoso fullname: Vizoso, Miguel email: m.vizoso.patino@gmail.com organization: Department of Molecular Pathology, Oncode Institute, Netherlands Cancer Institute – sequence: 2 givenname: Colin orcidid: 0000-0001-7848-9377 surname: E. J. Pritchard fullname: E. J. Pritchard, Colin organization: Mouse Clinic for Cancer and Aging, The Netherlands Cancer Institute – sequence: 3 givenname: Lorenzo surname: Bombardelli fullname: Bombardelli, Lorenzo organization: Division of Molecular Carcinogenesis and Oncode Institute, Netherlands Cancer Institute – sequence: 4 givenname: Bram orcidid: 0000-0003-1037-907X surname: van den Broek fullname: van den Broek, Bram organization: Cell Biophysics Group, Department of Cell Biology, The Netherlands Cancer Institute, BioImaging Facility, The Netherlands Cancer Institute – sequence: 5 givenname: Paul surname: Krimpenfort fullname: Krimpenfort, Paul organization: Mouse Clinic for Cancer and Aging, The Netherlands Cancer Institute – sequence: 6 givenname: Roderick L. orcidid: 0000-0003-0116-4130 surname: Beijersbergen fullname: Beijersbergen, Roderick L. organization: Division of Molecular Carcinogenesis and Oncode Institute, Netherlands Cancer Institute, NKI Robotics and Screening Center and Genomics Core Facility, The Netherlands Cancer Institute – sequence: 7 givenname: Kees orcidid: 0000-0001-7019-3440 surname: Jalink fullname: Jalink, Kees organization: Cell Biophysics Group, Department of Cell Biology, The Netherlands Cancer Institute, Swammerdam Institute for Life Sciences, University of Amsterdam – sequence: 8 givenname: Jacco orcidid: 0000-0001-8175-1647 surname: van Rheenen fullname: van Rheenen, Jacco email: j.v.rheenen@nki.nl organization: Department of Molecular Pathology, Oncode Institute, Netherlands Cancer Institute
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Cites_doi	10.1083/jcb.201907178 10.1038/ncb3509 10.1073/pnas.1701219114 10.1038/s41374-020-00482-5 10.1038/nmeth.3405 10.1016/j.stem.2015.02.006 10.15252/embj.2019102169 10.1016/j.ccr.2007.08.020 10.1038/nmeth.4222 10.1038/nmeth.2019 10.1038/s41467-020-16030-0 10.1093/bioinformatics/btx180 10.1038/ng.3905 10.1038/sj.gene.6364042 10.1038/nature19069 10.1038/nmeth.1524 10.1093/carcin/bgq191 10.7150/thno.47007 10.1016/j.celrep.2018.09.026 10.1038/nchembio.2063 10.1016/j.cell.2010.09.016 10.1038/s41592-019-0501-0 10.1039/D1SC01059J 10.1038/s41598-017-01684-6 10.1093/nar/gkm446 10.1073/pnas.1614057113 10.1038/ncomms7256 10.1038/s41467-019-08301-2 10.1038/nchembio.2205 10.1038/nbt1191 10.7554/eLife.61268 10.1109/ISBI.2016.7493463
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Snippet	The experimental need to engineer the genome both in time and space, has led to the development of several photoactivatable Cre recombinase systems. However,... Achieving spatial control of gene expression is important. Here the authors report an optimised photoactivatable Cre recombinase system, doxycycline- and...
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Title	A doxycycline- and light-inducible Cre recombinase mouse model for optogenetic genome editing
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