title: Continuous genetic recording with self-targeting CRISPR-Cas in human cells.
abstract: The ability to record molecular events in vivo would enable monitoring of signaling dynamics within cellular niches and critical factors that orchestrate cellular behavior. We present a self-contained analog memory device for longitudinal recording of molecular stimuli into DNA mutations in human cells. This device consists of a self-targeting guide RNA (stgRNA) that repeatedly directs Streptococcus pyogenes Cas9 nuclease activity toward the DNA that encodes the stgRNA, enabling localized, continuous DNA mutagenesis as a function of stgRNA expression. We demonstrate programmable and multiplexed memory storage in human cells triggered by exogenous inducers or inflammation, both in vitro and in vivo. This tool, Mammalian Synthetic Cellular Recorder Integrating Biological Events (mSCRIBE), provides a distinct strategy for investigating cell biology in vivo and enables continuous evolution of targeted DNA sequences.
authors.affiliation: Synthetic Biology Group, MIT Synthetic Biology Center, Massachusetts Institute of Technology (MIT), Cambridge, MA 02139, USA. Research Laboratory of Electronics, MIT, Cambridge, MA 02139, USA. Department of Electrical Engineering and Computer Science, MIT, Cambridge, MA 02139, USA.
authors.forename: Samuel D
authors.initials: SD
authors.lastname: Perli
authors.affiliation: Synthetic Biology Group, MIT Synthetic Biology Center, Massachusetts Institute of Technology (MIT), Cambridge, MA 02139, USA. Research Laboratory of Electronics, MIT, Cambridge, MA 02139, USA. Harvard-MIT Division of Health Sciences and Technology, Cambridge, MA 02139, USA.
authors.forename: Cheryl H
authors.initials: CH
authors.lastname: Cui
authors.affiliation: Synthetic Biology Group, MIT Synthetic Biology Center, Massachusetts Institute of Technology (MIT), Cambridge, MA 02139, USA. Research Laboratory of Electronics, MIT, Cambridge, MA 02139, USA. Department of Electrical Engineering and Computer Science, MIT, Cambridge, MA 02139, USA. Department of Biological Engineering, MIT, Cambridge, MA 02139, USA. timlu@mit.edu.
authors.forename: Timothy K
authors.initials: TK
authors.lastname: Lu
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D001483:Base Sequence / Q000235:genetics; D000076987:CRISPR-Associated Protein 9; D064113:CRISPR-Cas Systems; D058948:Cell Tracking / Q000379:methods*; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D004247:DNA / Q000235:genetics*; D004720:Endonucleases / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008568:Memory; D016296:Mutagenesis; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics / Q000378:metabolism*; D059010:Single-Cell Analysis / Q000379:methods*
chemical_list: D001426:Bacterial Proteins; D000094704:RNA, Guide, CRISPR-Cas Systems; D004247:DNA; D000076987:CRISPR-Associated Protein 9; C000606107:Cas9 endonuclease Streptococcus pyogenes; D004720:Endonucleases