Clonal analysis by tunable CRISPR-mediated excision


Journal article


Gilles, Schinko, Schacht, Enjolras, Averof
Development, vol. 146, 2019, pp. dev170969

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APA   Click to copy
Gilles, Schinko, Schacht, Enjolras, & Averof. (2019). Clonal analysis by tunable CRISPR-mediated excision. Development, 146, dev170969.


Chicago/Turabian   Click to copy
Gilles, Schinko, Schacht, Enjolras, and Averof. “Clonal Analysis by Tunable CRISPR-Mediated Excision.” Development 146 (2019): dev170969.


MLA   Click to copy
Gilles, et al. “Clonal Analysis by Tunable CRISPR-Mediated Excision.” Development, vol. 146, 2019, p. dev170969.


BibTeX   Click to copy

@article{gilles2019a,
  title = {Clonal analysis by tunable CRISPR-mediated excision},
  year = {2019},
  journal = {Development},
  pages = {dev170969},
  volume = {146},
  author = {Gilles and Schinko and Schacht and Enjolras and Averof}
}

Abstract

ABSTRACT Clonal marking techniques based on the Cre/lox and Flp/FRT systems are widely used in multicellular model organisms to mark individual cells and their progeny, in order to study their morphology, growth properties and developmental fates. The same tools can be adapted to introduce specific genetic changes in a subset of cells within the body, i.e. to perform mosaic genetic analysis. Marking and manipulating distinct cell clones requires control over the frequency of clone induction, which is sometimes difficult to achieve. Here, we present Valcyrie, a new method that replaces the conventional Cre or Flp recombinase-mediated excision of a marker cassette by CRISPR-mediated excision. A major advantage of this approach is that CRISPR efficiency can be tuned in a predictable fashion by manipulating the degree of sequence complementarity between the CRISPR guide RNA and its targets. We establish the method in the beetle Tribolium castaneum. We demonstrate that clone marking frequency can be tuned to generate embryos that carry single marked clones. The Valcyrie approach can be applied to a wide range of experimental settings, for example to modulate clone frequency with existing tools in established model organisms and to introduce clonal analysis in emerging experimental models. Summary: A clonal analysis approach based on CRISPR-mediated excision, which allows control over the frequency of clonal labelling, is proposed and tested in the beetle Tribolium castaneum.





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