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CMN Weekly (22 July 2022) - Your Weekly CRISPR Medicine News

Some of the best links we picked up around the internet

By: Gorm Palmgren - Jul. 22, 2022

Top picks

  • Danish researchers have shown that Cas12a nucleases can broadly utilise structurally engineered crRNAs with breaks or gaps in the otherwise highly-conserved loop region. The researchers tested 16 modifications ranging from engineered breaks (STAR-crRNAs) to large gaps (Gap-crRNAs), as well as nucleotide substitutions, and they all enabled gene-cutting in at least some of five Cas12a nucleases. The observed nuclease-specific differences in the crRNA tolerance may improve classification criteria and engineering strategies.
  • To treat autosomal dominant hearing loss, Chinese researchers have used cell sorting to precisely detect the efficiency of in vivo CRISPR-Cas9 gene editing in hair cells. Until now, it has been almost impossible to evaluate editing efficiencies because the whole cochlear tissue contains many non-target cells. In a mouse model, the authors show that a mutation in Kcnq4 responsible for hearing loss was repaired in 34% of hair cells, while the editing efficiency in the whole cochlear tissue was only 1.5%. Furthermore, gene editing improved hearing in treated animals as evaluated by several parameters.

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Detection

  • Chinese researchers propose a new Cas13a cascade cyclic signal enhancement strategy without amplification for detecting RNA viruses and disease-related RNAs. The method utilises intra-enzyme chain replacement-promoted Cas13a cascade cyclic reaction and is performed in 30 minutes with fluorometric readout. A detection limit of 75 aM was achieved with miRNA-21 as the target. Moreover, analyses of clinical respiratory syncytial virus (RSV) samples demonstrate good performance.
  • Another Chinese research team describes a CRISPR Cas12a-based detection system for miRNA-141. Cas12a is used to initiate a switchable ternary electrochemiluminescence (ECL) biosensor combined with a Co3O4@Au nanoemitter. In addition, a programmable enzyme-free hybrid chain reaction (HCR) amplifier on a paper working electrode (PWE) is used to assemble the nucleic acid tandem and accomplish the secondary recursion of the signal. As a result, the biosensor demonstrates high sensitivity and specificity, with a detection limit of 3.3 FM.
  • Researchers in India describe a CRISPR-Cas9-based lateral flow test for detecting SARS-CoV-2 RNA. It uses Cas9 from Francisella novicida and can detect specific nucleobase and nucleotide sequences. Detection depends upon the stoichiometric-based binding of FnCas9 ribonucleoprotein complex (RNP)-target sequences. The assay has been optimised to be conducted within one h and shows 100% sensitivity and 97% specificity in clinical samples across a range of viral loads.

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News: CMN Weekly (22 July 2022) - Your Weekly CRISPR Medicine News
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