2019
DOI: 10.1016/j.molcel.2019.10.024
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Dynamic Imaging of RNA in Living Cells by CRISPR-Cas13 Systems

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Cited by 271 publications
(300 citation statements)
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“…We also constructed dRfxCas13d-APEX2-NLS plasmid and designed the Rfx-crRNAs targeting NEAT1 to study paraspeckles. We found that the localization of dRfxCas13d-APEX2-NLS had no difference between the non-targeting crRNA group and the NEAT1 targeting crRNA group (data not show), which is consistent with results of Chen lab [34] . These data suggested that dRfxCas13d is not suitable for CBRPP to study RNA-protein interactions.…”
Section: Resultssupporting
confidence: 91%
“…We also constructed dRfxCas13d-APEX2-NLS plasmid and designed the Rfx-crRNAs targeting NEAT1 to study paraspeckles. We found that the localization of dRfxCas13d-APEX2-NLS had no difference between the non-targeting crRNA group and the NEAT1 targeting crRNA group (data not show), which is consistent with results of Chen lab [34] . These data suggested that dRfxCas13d is not suitable for CBRPP to study RNA-protein interactions.…”
Section: Resultssupporting
confidence: 91%
“…Here, we have repurposed the RNA-guided RNA endonuclease activity of Cas13d in mammalian cells against emergent viral targets, SARS-CoV-2 and IAV, in our PAC-MAN strategy. This expands the applications of CRISPR-Cas13 systems in addition to their uses for diagnostics such as SHERLOCK and live-cell imaging [30][31][32][33] . Prior work showed Cas13a/b systems could inhibit ssRNA viruses including influenza, which is consistent with our observed high efficiency of SARS-CoV-2 reporter and IAV inhibition using the PAC-MAN approach 31 .…”
Section: Discussion and Perspectivesmentioning
confidence: 95%
“…(4)(5)(6) Currently, the most ubiquitous approach for visualizing mRNA uses GFP-fused RNA binding proteins such as MS2, λN, PCP, or Cas proteins. (7)(8)(9)(10) These fluorescently-tagged proteins recognize a specific sequence that is incorporated multiple times onto ROI. While this does enable visualization of RNAs in living cells, these methodologies suffer from the fact that the unbound fluorescent protein creates significant background signal.…”
mentioning
confidence: 99%