CRISPR-Cas9: Difference between revisions
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<StructureSection load='4qyz' size='450' side='right' scene='74/742625/Cv/4' caption=''> | <StructureSection load='4qyz' size='450' side='right' scene='74/742625/Cv/4' caption=''> | ||
==Background== | |||
Highlights | |||
*CRISPR-Cas9 is a powerful tool to modulate transcription in wide range of cell types. | |||
*An expanding set of CRISPR-based transcription effectors is available. | |||
*Gene networks can be efficiently probed and modified for biotechnology applications.<ref name="Did">PMID:27344519</ref> | |||
CRISPR-Cas9 has recently emerged as a promising system for multiplexed genome editing as well as epigenome and transcriptome perturbation. Due to its specificity, ease of use and highly modular programmable nature, it has been widely adopted for a variety of applications such as genome editing, transcriptional inhibition and activation, genetic screening, DNA localization imaging, and many more. In this review, we will discuss non-editing applications of CRISPR-Cas9 for transcriptome perturbation, metabolic engineering, and synthetic biology.<ref name="Did">PMID:27344519</ref> | |||
==Crystal structure of a CRISPR RNA-guided surveillance complex, Cascade, bound to a ssDNA target<ref>PMID:25123481</ref>== | ==Crystal structure of a CRISPR RNA-guided surveillance complex, Cascade, bound to a ssDNA target<ref>PMID:25123481</ref>== | ||
The <scene name='74/742625/Cv/5'>crystal structure of ssDNA-bound Cascade has the seahorse architecture</scene>. The body is formed by a helical filament of six Cas7 subunits (Cas7.1 to 7.6) wrapped around the crRNA guide, with a head-to-tail dimer of Cse2 (Cse2.1 and Cse2.2) at the belly. Cas6e and the 3′ handle of crRNA cap the Cas7 filament at the head while Cas5 and the 5′ handle cap the tail. The N-terminal base of Cse1 is positioned at the tail of the filament; the C-terminal four-helix bundle contacts Cse2.2. The ssDNA target is juxtaposed to the guide region of the crRNA in a groove formed by the Cas7 filament, the four-helix bundle of Cse1, and the Cse2 dimer. | The <scene name='74/742625/Cv/5'>crystal structure of ssDNA-bound Cascade has the seahorse architecture</scene>. The body is formed by a helical filament of six Cas7 subunits (Cas7.1 to 7.6) wrapped around the crRNA guide, with a head-to-tail dimer of Cse2 (Cse2.1 and Cse2.2) at the belly. Cas6e and the 3′ handle of crRNA cap the Cas7 filament at the head while Cas5 and the 5′ handle cap the tail. The N-terminal base of Cse1 is positioned at the tail of the filament; the C-terminal four-helix bundle contacts Cse2.2. The ssDNA target is juxtaposed to the guide region of the crRNA in a groove formed by the Cas7 filament, the four-helix bundle of Cse1, and the Cse2 dimer. | ||