CRISPR/Cas12a collateral cleavage activity for an ultrasensitive assay of RNase H

2022 ◽  
Author(s):  
Hansol Kim ◽  
Seoyoung Lee ◽  
Jinhwan Lee ◽  
Hyun Gyu Park

We herein describe an ultrasensitive RNase H assay by utilizing CRISPR/Cas12a collateral cleavage activity. In this technique, a chimeric duplex probe consisting of activator DNA and its complementary blocker RNA...

2013 ◽  
Vol 455 (2) ◽  
pp. 179-184 ◽  
Author(s):  
Brian D. Herman ◽  
Nicolas Sluis-Cremer

Pre-steady-state kinetics were used to define the mechanisms by which efavirenz and a β-thujaplicinol analogue modulate the RNase H activity of HIV-1 reverse transcriptase. Both inhibitors do not affect polymerase-dependent cleavages, but significantly affect the rates of polymerase-independent cleavages.


1990 ◽  
Vol 55 (11) ◽  
pp. 2781-2786 ◽  
Author(s):  
Valerii G. Metelev ◽  
Natalya F. Krynetskaya ◽  
Andrei A. Purmal ◽  
Zoya A. Shabarova ◽  
Zdeněk Točík ◽  
...  

Undecanucleotide dA-dC-dC-dA-Cm-Cm-dG-Cm-dG-Cm-dT (Cm = 2'-O-methylcytidine) was prepared and used as a probe for splitting of 5S rRNA fragment by RNase H (E. coli). It was found that substitution of 2'-deoxycytidine of d(ACCACCGCGCT) by 2'-O-methylcytidine in positions 5, 6, 8 and 10 provided regiospecific splitting of RNA fragment by RNase H in the presence of modified probe between U25 and C26.


2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Wenjing Qi ◽  
Erika D. V. Gromoff ◽  
Fan Xu ◽  
Qian Zhao ◽  
Wei Yang ◽  
...  

AbstractMulticellular organisms coordinate tissue specific responses to environmental information via both cell-autonomous and non-autonomous mechanisms. In addition to secreted ligands, recent reports implicated release of small RNAs in regulating gene expression across tissue boundaries. Here, we show that the conserved poly-U specific endoribonuclease ENDU-2 in C. elegans is secreted from the soma and taken-up by the germline to ensure germline immortality at elevated temperature. ENDU-2 binds to mature mRNAs and negatively regulates mRNA abundance both in the soma and the germline. While ENDU-2 promotes RNA decay in the soma directly via its endoribonuclease activity, ENDU-2 prevents misexpression of soma-specific genes in the germline and preserves germline immortality independent of its RNA-cleavage activity. In summary, our results suggest that the secreted RNase ENDU-2 regulates gene expression across tissue boundaries in response to temperature alterations and contributes to maintenance of stem cell immortality, probably via retaining a stem cell specific program of gene expression.


2021 ◽  
Vol 178 ◽  
pp. 113019
Author(s):  
Kai Zhang ◽  
Zhenqiang Fan ◽  
Bo Yao ◽  
Yuedi Ding ◽  
Jing Zhao ◽  
...  
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