Endogenous CRISPR/Cas9 arrays for scalable whole-organism lineage tracing
Mostra el registre complet Registre parcial de l'ítem
- dc.contributor.author Cotterell, James
- dc.contributor.author Vila-Cejudo, Marta
- dc.contributor.author Batlle Morera, Laura
- dc.contributor.author Sharpe, James
- dc.date.accessioned 2020-10-08T06:21:07Z
- dc.date.available 2020-10-08T06:21:07Z
- dc.date.issued 2020
- dc.description.abstract The past decade has seen a renewed appreciation of the central importance of cellular lineages to many questions in biology (especially organogenesis, stem cells and tumor biology). This has been driven in part by a renaissance in genetic clonal-labeling techniques. Recent approaches are based on accelerated mutation of DNA sequences, which can then be sequenced from individual cells to re-create a 'phylogenetic' tree of cell lineage. However, current approaches depend on making transgenic alterations to the genome in question, which limit their application. Here, we introduce a new method that completely avoids the need for prior genetic engineering, by identifying endogenous CRISPR/Cas9 target arrays suitable for lineage analysis. In both mouse and zebrafish, we identify the highest quality compact arrays as judged by equal base composition, 5' G sequence, minimal likelihood of residing in the functional genome, minimal off targets and ease of amplification. We validate multiple high-quality endogenous CRISPR/Cas9 arrays, demonstrating their utility for lineage tracing. Our pragmatically scalable technique thus can produce deep and broad lineages in vivo, while removing the dependence on genetic engineering.
- dc.description.sponsorship This work was funded by the core program funding of the European Molecular Biology Laboratory with support from the European Research Council through SIMBIONT (670555). Deposited in PMC for immediate release
- dc.format.mimetype application/pdf
- dc.identifier.citation Cotterell J, Vila-Cejudo M, Batlle-Morera L, Sharpe J. Endogenous CRISPR/Cas9 arrays for scalable whole-organism lineage tracing. Development. 2020 May 12; 147(9): dev184481. DOI: 10.1242/dev.184481
- dc.identifier.doi http://dx.doi.org/10.1242/dev.184481
- dc.identifier.issn 0950-1991
- dc.identifier.uri http://hdl.handle.net/10230/45429
- dc.language.iso eng
- dc.publisher Company of Biologists
- dc.relation.ispartof Development. 2020 May 12;147(9):dev184481
- dc.relation.projectID info:eu-repo/grantAgreement/EC/H2020/670555
- dc.rights © 2020. Published by The Company of Biologists Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed
- dc.rights.accessRights info:eu-repo/semantics/openAccess
- dc.rights.uri https://creativecommons.org/licenses/by/4.0
- dc.subject.other Llinatge cel·lular
- dc.subject.other Morfogènesi
- dc.subject.other Cèl·lules mare
- dc.subject.other Seqüència de nucleòtids
- dc.subject.other Genomes
- dc.title Endogenous CRISPR/Cas9 arrays for scalable whole-organism lineage tracing
- dc.type info:eu-repo/semantics/article
- dc.type.version info:eu-repo/semantics/publishedVersion