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Cell‐Lineage Tracing in Zebrafish Embryos with an Expanded Genetic Code by Wes Brown

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From the article: Cell‐lineage tracing is used to study embryo development and stem‐cell differentiation as well as to document tumor cell heterogeneity. Cre recombinase‐mediated cell labeling is the preferred approach; however, its utility is restricted by when and where DNA recombination takes place. We generated a photoactivatable Cre recombinase by replacing a critical residue in its active site with a photocaged lysine derivative through genetic code expansion in zebrafish embryos. This allows high spatiotemporal control of DNA recombination by using 405 nm irradiation. Importantly, no background activity is seen before irradiation, and, after light‐triggered removal of the caging group, Cre recombinase activity is restored. We demonstrate the utility of this tool as a cell‐lineage tracer through its activation in different regions and at different time points in the early embryo. Direct control of Cre recombinase by light will allow more precise DNA recombination, thereby enabling more nuanced studies of metazoan development and disease.

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