+Search query
-Structure paper
Title | Structural basis of target DNA recognition by CRISPR-Cas12k for RNA-guided DNA transposition. |
---|---|
Journal, issue, pages | Mol Cell, Vol. 81, Issue 21, Page 4457-44466.e5, Year 2021 |
Publish date | Nov 4, 2021 |
![]() | Renjian Xiao / Shukun Wang / Ruijie Han / Zhuang Li / Clinton Gabel / Indranil Arun Mukherjee / Leifu Chang / ![]() |
PubMed Abstract | The type V-K CRISPR-Cas system, featured by Cas12k effector with a naturally inactivated RuvC domain and associated with Tn7-like transposon for RNA-guided DNA transposition, is a promising tool for ...The type V-K CRISPR-Cas system, featured by Cas12k effector with a naturally inactivated RuvC domain and associated with Tn7-like transposon for RNA-guided DNA transposition, is a promising tool for precise DNA insertion. To reveal the mechanism underlying target DNA recognition, we determined a cryoelectron microscopy (cryo-EM) structure of Cas12k from cyanobacteria Scytonema hofmanni in complex with a single guide RNA (sgRNA) and a double-stranded target DNA. Coupled with mutagenesis and in vitro DNA transposition assay, our results revealed mechanisms for the recognition of the GGTT protospacer adjacent motif (PAM) sequence and the structural elements of Cas12k critical for RNA-guided DNA transposition. These structural and mechanistic insights should aid in the development of type V-K CRISPR-transposon systems as tools for genome editing. |
![]() | ![]() ![]() ![]() |
Methods | EM (single particle) |
Resolution | 3.65 - 3.8 Å |
Structure data | EMDB-24142, PDB-7n3o: EMDB-24143, PDB-7n3p: |
Source |
|
![]() | RNA BINDING PROTEIN/RNA / CRISPR Cas / RNA binding protein / protein-RNA complex / RNA BINDING PROTEIN-RNA complex / RNA BINDING PROTEIN/RNA/DNA / protein-RNA-DNA complex / RNA BINDING PROTEIN-RNA-DNA complex |