Homing guide RNAs that target Cas9 to their own loci generate diverse barcodes that can trace the lineage of cells.
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In this study, the scientists present an approach for engineering evolving DNA barcodes in living cells. A homing guide RNA (hgRNA) scaffold directs the Cas9–hgRNA complex to the DNA locus of the hgRNA itself. This integrated approach will have wide-ranging applications, such as in deep lineage tracing, cellular barcoding, molecular recording, dissecting cancer biology, and connectome mapping.
Targeting point mutations using CRISPR/Cas9 so far has required efficient homologous recombination (HR) and donor oligonucleotides. In a recent Nature paper, Komor and colleagues (2016) describe a way to make specific base changes that does not depend on HR or donor DNA and does not involve making double-strand breaks.
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In this study, the scientists present an approach for engineering evolving DNA barcodes in living cells. A homing guide RNA (hgRNA) scaffold directs the Cas9–hgRNA complex to the DNA locus of the hgRNA itself. This integrated approach will have wide-ranging applications, such as in deep lineage tracing, cellular barcoding, molecular recording, dissecting cancer biology, and connectome mapping.