Here, we demonstrate that CRISPR-Cas13 systems can be repurposed to assist trans-splicing of exogenous RNA fragments into an endogenous pre-mRNA transcript, a method termed CRISPR Assisted mRNA Fragment Trans-s
Here, we demonstrate that CRISPR- Cas13 systems can be repurposed to assist trans-splicing of exogenous RNA fragments into an endogenous pre-mRNA transcript, a method termed CRISPR Assisted mRNA Fragment Trans-splicing (CRAFT). Using split reporter-based assays, we evaluate orthogonal Cas13 ...
Conventional CRISPR-assisted biosensing was not applicable for diagnosing methylation at individual CpG sites. Dongen et al. designed an in vitro diagnostic method based on CRISPR/Cas12a. By using methylation-sensitive restriction endonucleases (MSREs), the digestion of unmethylated fragments was facilitate...
2f) might be also plasmid insertions or due to an inefficient intron removal during the pre-mRNA splicing, that can occur in experiments conducted with a transfected plasmid reporter75. Unwanted outcomes of DSB repair after canonical CRISPR/Cas9 targeting are a potential threat even if occurring ...
gene expression and function [225]. Targeted fragment deletion can also be used to delete mis-spliced exons that contained stop codons and were resulted from mutations to splicing elements, such as cis -regulatory elements, core spliceosomal components and trans -acting regulatory factors.For example...
The reduced RNA-cleavage efficiency and the likelihood for off-target effects on host DNA have meant that RNA-targeting orthologs of Cas9 are unlikely to be useful in targeting RNA transcripts directly, such as non-coding RNA (ncRNA), messenger RNA (mRNA), or viral RNA genomes.77,78 Moreove...
The resulting 821 bp DNA fragment (Figure S1A) was synthesized and inserted into the SacII and BstZ17I sites of p85Cas9. To modify pCas9gRNA-cpaAIR for genome editing via deletion of cpaAIR, splicing by overlap extension (SOE) PCR was utilized to fuse 1,028 bp and 1,057 bp ...
endogenous tRNA system as a robust reagent can intensify the genome-editing capability and efficiency without requiring additional nucleases or introducing RNAs apart from Cpf1/crRNA (Fig.4). In addition, intronic PTG (inPTG) uses the endogenous mRNA splicing and tRNA processing machineries to ...
mRNA decay (NMD), thereby degrading mRNA and terminating protein translation [48]. However, BEs do not directly interfere with splicing processes like some newer technologies, such as CRISPR-mediated RNA trans-splicing, which may provide a more immediate approach to controlling RNA splicing [49]....
CRISPR-Cas12a-based nucleic acid amplification-free DNA biosensor via au nanoparticle-assisted metal-enhanced fluorescence and colorimetric analysis. Nano Lett. 2021;21:693–9. https://doi.org/10.1021/acs.nanolett.0c04303. Article CAS PubMed Google Scholar Wang J, Gui C, Zhu J, Zhu B, ...