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We began analyzing https://pmc.ncbi.nlm.nih.gov/articles/PMC5694763/, but it redirected us to https://pmc.ncbi.nlm.nih.gov/articles/PMC5694763/. The analysis below is for the second page.
Title[redir]:
Mapping the sugar dependency for rational generation of a DNA-RNA hybrid-guided Cas9 endonuclease - PMC
Description:
The CRISPR–Cas9 RNA-guided endonuclease system allows precise and efficient modification of complex genomes and is continuously developed to enhance specificity, alter targeting and add new functional moieties. However, one area yet to be explored ...
Title[redir]:
Mapping the sugar dependency for rational generation of a DNA-RNA hybrid-guided Cas9 endonuclease - PMC
Description:
The CRISPR–Cas9 RNA-guided endonuclease system allows precise and efficient modification of complex genomes and is continuously developed to enhance specificity, alter targeting and add new functional moieties. However, one area yet to be explored ...
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Keywords {🔍}
dna, cas, activity, fig, doi, target, binding, rna, crrna, tracr, complex, molecules, supplementary, cleavage, crhyb, molecule, nuclease, crxna, pubmed, google, scholar, pmc, tracrrna, guide, article, sequence, assay, fret, free, reduced, residues, protein, complexes, vitro, hybrid, cells, buffer, region, generated, aavs, data, specificity, cellular, showed, hyb, crhybs, assays, show, design, crdna,
Topics {✒️}
n-terminal 6×hn tag 100 µm isopropyl β-d-1-thiogalactopyranoside high-fidelity crispr–cas9 nucleases biotin–streptavidin interactions time-binned histogram shows time-binned fret histogram fully rna-free endonucleases rcuk mc-a658-5ty10 activation-induced deoxycytidine deaminase double-sided sticky tape time-binned fret histograms rna-guided cas9 endonuclease rna-guided endonuclease cas9 crispr-based genome editing author contributions engineered crispr-cas9 nucleases hybrid dna–rna crispr dynamic mid-fret state pmc beta search dna–rna hybrid strand29 double-strand break caused single-molecule results suggest weak target-binding affinity c2′-endo sugar pucker single-molecule fret assay active high-fret conformation cas9 remains target-bound stable high-fret state recent single-molecule studies split-cas9 enzyme complex stable high-fret conformation previously unobserved low-fret 5′-biotin crhyb-binding defect arises 790 bp double-stranded dna stable high-fret trajectories dna–rna hybrid molecules single base-pair mismatch hybrid dna–rna crxna xna-programmable endonuclease low-protein-binding tubes cas9–target interaction increases double-stranded rnas reduced target-binding affinity dna–dna duplex destabilising agarose gel electrophoresis rna-guided nucleases direct specific cas9 ribose-free crxna specific unproductive binding
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