Synthego Corporation, 3565 Haven Avenue, Menlo Park, CA, 94025, USA.
Freenome, 259 East Grand Ave, South San Francisco, CA, 94080, USA.
Nat Commun. 2020 Oct 7;11(1):5041. 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.
将 CRISPR-Cas9 组件引入细胞后，基因组编辑会持续进行，直到细胞过程降解其组件核酸和蛋白质。这种不受控制的反应可能会导致意想不到的后果，包括脱靶编辑和染色体易位。为了解决这个问题，我们开发了一种称为 CRISPRoff 的 sgRNA 光诱导降解方法。在这里，我们表明，核糖核蛋白的光诱导失活可减弱细胞内的基因组编辑，并通过选择性照明实现可滴定的编辑效率和空间模式。
