CRISPRoff enables spatio-temporal control of CRISPR editing

• Published in: Nature communications Vol. 11; no. 1; pp. 5041 - 7
• Main Authors: Carlson-Stevermer, Jared, Kelso, Reed, Kadina, Anastasia, Joshi, Sahil, Rossi, Nicholas, Walker, John, Stoner, Rich, Maures, Travis
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 07.10.2020; Nature Portfolio
• Subjects: 13/106; 13/56; 42/109; 45/23; 45/77; 631/1647/1511; 631/208/4041/3196; 631/92/2783; Cell Line, Tumor; CRISPR-Cas Systems - genetics; CRISPR-Cas Systems - radiation effects; DNA Breaks, Double-Stranded; Feasibility Studies; Gene Editing - methods; HEK293 Cells; Humanities and Social Sciences; Humans; Light; multidisciplinary; Ribonucleoproteins - metabolism; RNA Stability - radiation effects; RNA, Guide, CRISPR-Cas Systems - metabolism; RNA, Guide, CRISPR-Cas Systems - radiation effects; Science; Science (multidisciplinary); Translocation, Genetic
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-020-18853-3

Following introduction of CRISPR-Cas9 components into a cell, genome editing occurs unabated until degradation of its component nucleic acids and proteins by cellular processes. This uncontrolled reaction can lead to unintended consequences including off-target editing and chromosomal translocations. To address this, we develop a method for light-induced degradation of sgRNA termed CRISPRoff. Here we show that light-induced inactivation of ribonucleoprotein attenuates genome editing within cells and allows for titratable levels of editing efficiency and spatial patterning via selective illumination. Uncontrolled gene editing can lead to off-target effects and chromosomal translocations. Here the authors develop CRISPRoff, light degradable sgRNAs for titratable and spatially defined gene editing.