Tag: RfxCas13d

Intrinsic targeting of host RNA by Cas13 constrains its utility

Abudayyeh, O. O. et al. C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector. Science 353, aaf5573 (2016). Article  PubMed  PubMed Central  Google Scholar  Terns, M. P. CRISPR-based technologies: impact of RNA-targeting systems. Mol. Cell 72, 404–412 (2018). Article  CAS  PubMed  PubMed Central  Google Scholar  Abudayyeh, O. O. et al….

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Mitigating a TDP-43 proteinopathy by concentrating on ataxin-2 utilizing

Concentrating on ataxin-2 with RfxCas13d Due to its potential to affect TDP-43-associated toxicity20,25,26, ataxin-2 has emerged as a doubtlessly broadly relevant goal for ALS-FTD, as TDP-43 pathology is noticed in ~97% of ALS instances1,4,5 and ~45% FTD occurrences1,2,4. Given this, we sought to find out if RNA-targeting CRISPR effector proteins…

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Precision Targeting of Na-K-2Cl cotransporter (NKCC1) RNA In Vitro with RfxCas13d

Abstract The Na-K-2Cl cotransporter (NKCC1) is considered an attractive drug target in the Central nervous system (CNS) for treating various CNS disorders. However, the specific role of NKCC1 in different types following injury within the CNS is not well understood due to its expression in multiple cell types. Additionally, there…

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Stable expression of large transgenes via the knock-in of an integrase-deficient lentivirus

CLIP enables high-efficiency knock-in into an essential locus To create CLIP, we leverage integrase-deficient lentiviruses (IDLV) to deliver an HDR donor template as a viral RNA genome to cells25,26. IDLV consists of lentiviral components but with a D64V mutation in the integrase, maintaining this enzyme’s ability to package and nuclear…

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Collateral activity of the CRISPR/RfxCas13d system in human cells

Cell culture HEK293T, MEF-1, and U87 cell lines were used in this research. HEK293T was purchased from ATCC. MEF-1, and U87 cell lines were gifts from Muredach P. Reilly, Peter A. Sims, respectively. HEK293T, MEF-1, and U87 cells were cultured in DMEM with 4.5 g/L D-Glucose, supplemented with 10% fetal bovine…

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The era of cryptic exons: implications for ALS-FTD | Molecular Neurodegeneration

‘TDP-43 proteinopathy’ refers to human diseases pathologically defined by cellular nuclear-to-cytoplasmic mislocalisation and aggregation of TAR DNA-binding protein-43 (TDP-43), an RNA-binding protein with a crucial role in RNA metabolism and splicing. TDP-43 proteinopathy encompasses a wide spectrum of progressive neurodegenerative diseases and phenotypes. It is a key feature in ~ 97%…

Continue Reading The era of cryptic exons: implications for ALS-FTD | Molecular Neurodegeneration

The collateral activity of RfxCas13d can induce lethality in a RfxCas13d knock-in mouse model

Background: The CRISPR-Cas13 system is an RNA-guided RNA-targeting system and has been widely used in transcriptome engineering with potentially important clinical applications. However, it is still controversial whether Cas13 exhibits collateral activity in mammalian cells. Results: Here, we find that knocking down gene expression using RfxCas13d in the adult brain…

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Collateral cleavage of 28s rRNA by RfxCas13d causes death of mice

Abstract Summary The CRISPR-Cas13 system is an RNA-guided RNA-targeting system, and has been widely used in transcriptome engineering with potentially important clinical applications. However, it is still controversial whether Cas13 exhibits collateral activity in mammalian cells. Here, we found that knocking down gene expression using RfxCas13d in the adult brain…

Continue Reading Collateral cleavage of 28s rRNA by RfxCas13d causes death of mice