2022
DOI: 10.1016/j.bios.2022.114074
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Live imaging of RNA and RNA splicing in mammalian cells via the dcas13a-SunTag-BiFC system

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Cited by 13 publications
(8 citation statements)
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“…Cas13a is covalently linked to multiple tandem GCN4 peptides and recruits multiple FPs coupled with scFv to target RNA sites. Furthermore, combining the BiFc (bimolecular fluorescence complementation) system with Cas13a-SunTag system, which relies on split FPs, has showed a better signal-to-noise ratio than that of the MS2-MCP system (Figure 3c[i], lower panel; M. Chen et al, 2022).…”
Section: Rna Tracking and Imagingmentioning
confidence: 99%
“…Cas13a is covalently linked to multiple tandem GCN4 peptides and recruits multiple FPs coupled with scFv to target RNA sites. Furthermore, combining the BiFc (bimolecular fluorescence complementation) system with Cas13a-SunTag system, which relies on split FPs, has showed a better signal-to-noise ratio than that of the MS2-MCP system (Figure 3c[i], lower panel; M. Chen et al, 2022).…”
Section: Rna Tracking and Imagingmentioning
confidence: 99%
“…The dCas9-Suntag system has been used for low-repetitive and non-repetitive genomic loci, which increases the fluorescent signal-to-background ratio (S/B) in comparison with the dCas9-EGFP system ( Shao et al, 2018b ). Additionally, the dCas13-Suntag system has also been used to label RNA based on the same amplification principle ( Xu et al, 2021 ; Chen et al, 2022 ). The sgRNA has been redesigned to insert two or more copies of MS2 or PP7 for MCP-FPs or PCP-FPs binding in CRISPR-based labeling systems ( Fu et al, 2016 ; Shao et al, 2016 ; Wang et al, 2016 ).…”
Section: Fluorescent Signal Amplification Techniquesmentioning
confidence: 99%
“…Therefore, efficient delivery or optimization of sgRNA structure is a key aspect worth considering in both imaging and editing. To enhance the specificity and reduce background noise in nucleic acid imaging within living cells, it is crucial to establish a precise match between the CRISPR probe and the targeted genomic or RNA loci (Chen et al, 2022). This may necessitate ongoing optimization with diverse guided-RNA sequences to attain the desired level of specificity.…”
Section: Target Accessibility and Cas Effector/grna Ratiomentioning
confidence: 99%