Aptazyme-embedded guide RNAs enable ligand-responsive genome editing and transcriptional activation

• Published in: Nature communications Vol. 8; no. 1; pp. 15939 - 8
• Main Authors: Tang, Weixin, Hu, Johnny H., Liu, David R.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 28.06.2017; Nature Publishing Group; Nature Portfolio
• Subjects: 45/41; 631/1647/1511; 631/61/201/2110; Active control; Canonical forms; Catalysis; CRISPR; CRISPR-Cas Systems; Editing; Gene Editing; Gene expression; Gene regulation; Genome editing; Genomes; HEK293 Cells; Humanities and Social Sciences; Humans; Ligands; Mammalian cells; multidisciplinary; Nuclease; Nucleic Acid Conformation; Nucleotide sequence; RNA, Catalytic - genetics; RNA, Catalytic - metabolism; RNA, Guide, CRISPR-Cas Systems - chemistry; RNA, Guide, CRISPR-Cas Systems - genetics; RNA, Guide, CRISPR-Cas Systems - metabolism; Science; Science (multidisciplinary); Transcription activation; Transcriptional Activation
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/ncomms15939

Programmable sequence-specific genome editing agents such as CRISPR-Cas9 have greatly advanced our ability to manipulate the human genome. Although canonical forms of genome-editing agents and programmable transcriptional regulators are constitutively active, precise temporal and spatial control over genome editing and transcriptional regulation activities would enable the more selective and potentially safer use of these powerful technologies. Here, by incorporating ligand-responsive self-cleaving catalytic RNAs (aptazymes) into guide RNAs, we developed a set of aptazyme-embedded guide RNAs that enable small molecule-controlled nuclease-mediated genome editing and small molecule-controlled base editing, as well as small molecule-dependent transcriptional activation in mammalian cells. CRISPR-Cas9 is a powerful technique for manipulating the human genome, however temporal and spatial control of activity would facilitate safer, more selective use. Here the authors incorporate aptazymes into guide RNAs to allow small molecule activation of CRISPR-Cas9.