Abstract
We present here an approach for engineering evolving DNA barcodes in living cells. The methodology entails use of a homing guide RNA (hgRNA) scaffold that directs the Cas9-hgRNA complex to target the DNA locus of the hgRNA itself. We show this homing CRISPR-Cas9 system acts as an expressed evolving genetic barcode, and corresponding small RNAs can be assayed as single molecules in situ. This integrated approach will have wide ranging applications, such as in deep lineage tracing, cellular barcoding, molecular recording, dissecting cancer biology, and connectome mapping.
Copyright
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
Posted July 26, 2016.
