We have developed a novel live-cell genomic imaging platform, termed CRISPR/MB, which comprises dCas9, an MB, and an sgRNA engineered with an MB target sequence in the stem–loop 2 region, and demonstrated its capacity for quantitative, dynamic and dual-color analysis of genomic loci in human cells.
We have developed self-assembled RNA nanostructures that can inhibit HIV-1 virus assembly, achieved through hybridization of multiple artificial small RNAs with a stem–loop structure (STL) that we identify as a prominent ligand of Gag that can inhibit virus particle production via STL-Gag interactions.
In contrast to the general notion that vRNA only triggers Gag assembly and is dispensable for subsequent assembly, we found that vRNA is indispensable throughout assembly, scaffolding the formation of assembly intermediates and maintaining their architectures via balancing of external forces acting on the assembly environment.
Article-Genomic Labeling
Our results indicated that false-positive foci do not result from nonspecific accumulation of sgRNAs as previously speculated. Instead, they pointed to a previously undescribed role of cryptic plasmid transcripts, which arise from spurious transcriptional activities occurring within plasmid DNA regions other than the intended promoter-gene cassette (i.e. transcription unit). These unintended transcripts are sequestered by paraspeckle proteins FUS, SFPQ and PSPC1 rather than accumulating around their producer plasmids, leading to false-positive foci formation. Based on these findings, we presented an easy-to-adapt strategy to minimize the generation of false-positive signals. Read More ›
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