Chromatin conformation,localization,and dynamics are crucial regulators of cellular behaviors. Although fluorescence in situ hybridization-based techniques have been widely utilized for investigating chromatin archite...Chromatin conformation,localization,and dynamics are crucial regulators of cellular behaviors. Although fluorescence in situ hybridization-based techniques have been widely utilized for investigating chromatin architectures in healthy and diseased states,the requirement for cell fix-ation precludes the comprehensive dynamic analysis necessary to fully understand chromatin activ-ities. This has spurred the development and application of a variety of imaging methodologies for visualizing single chromosomal loci in the native cellular context. In this review,we describe currently-available approaches for imaging single genomic loci in cells,with special focus on clus-tered regularly interspaced short palindromic repeats (CRISPR)-based imaging approaches. In addition,we discuss some of the challenges that limit the application of CRISPR-based genomic imaging approaches,and potential solutions to address these challenges. We anticipate that,with continued refinement of CRISPR-based imaging techniques,significant understanding can be gained to help decipher chromatin activities and their relevance to cellular physiology and pathogenesis.展开更多
With numerous advancements in novel biochemical techniques, our knowledge of the role of RNAs in the regulation of cellular physiology and pathology has grown significantly over the past several decades. Nevertheless,...With numerous advancements in novel biochemical techniques, our knowledge of the role of RNAs in the regulation of cellular physiology and pathology has grown significantly over the past several decades. Nevertheless, detailed information regarding RNA processing, trafficking, and localization in living cells has been lacking due to technical limitations in imaging single RNA transcripts in living cells with high spatial and temporal resolution. In this review, we discuss tech- niques that have shown great promise for single RNA imaging, followed by highlights in our recent work in the development of molecular beacons （MBs）, a class of nanoscale oligonucleotide-probes, for detecting individual RNA transcripts in living cells. With further refinement of MB design and development of more sophisticated fluorescence microscopy techniques, we envision that MB-based approaches could promote new discoveries of RNA functions and activities.展开更多
We assessed expression of IL-20 and its receptors in psoriasis,given the recent implication of IL-20 in epidermal hyperplasia.Psoriatic lesional(LS)skin consistently expressed more IL-20 mRNA than nonlesional(NL)skin....We assessed expression of IL-20 and its receptors in psoriasis,given the recent implication of IL-20 in epidermal hyperplasia.Psoriatic lesional(LS)skin consistently expressed more IL-20 mRNA than nonlesional(NL)skin.Immunoreactivity to IL-20 protein was greater in LS tissue and mainly localized to infiltrating CD68+/CD11c+(myeloid-derived)dermal leukocytes.Because this contrasted with earlier reports of a keratinocyte source,we assessed IL-20 mRNA expression in a variety of cells in vitro,and confirmed a myeloid-derived cellular source(monocytes).Plastic adhesion,activation of β2 integrins,and incubation with tumor necrosis factor-αstimulated expression in these cells.IL-20 receptor(IL-20R)αand IL-20RβmRNA was decreased in LS versus NL skin,which also contrasted with earlier findings.To investigate the relationship between IL-20 and disease activity,we examined psoriasis patients treated with the CD2-targeted agent alefacept.In therapeutic responders,lesional IL-20 mRNA decreased to NL levels,suggesting that CD2+leukocytes may proximally regulate IL-20.Finally,to assess IL-20 function,we used microarrays to screen IL-20-treated keratinocytes,which demonstrated upregulation of disease-related and IFN-γ-induced genes.Hence,IL-20 may influence inflammation through IFN-like effects.Together,these data indicate that IL-20 may be an important effector cytokine in psoriasis,and that its inhibition may represent a potential therapeutic target.展开更多
基金supported by grants from the National Key R&D Program of China (Grant Nos. 2016YFA0501603 and 2016YFA0100702)National Natural Science Foundation of China (Grant No. 31771583)+1 种基金Beijing Natural Science Foundation (Grant No. 7162114)Beijing Municipal R&D Key Project (Grant No. Z151100003915081), China
文摘Chromatin conformation,localization,and dynamics are crucial regulators of cellular behaviors. Although fluorescence in situ hybridization-based techniques have been widely utilized for investigating chromatin architectures in healthy and diseased states,the requirement for cell fix-ation precludes the comprehensive dynamic analysis necessary to fully understand chromatin activ-ities. This has spurred the development and application of a variety of imaging methodologies for visualizing single chromosomal loci in the native cellular context. In this review,we describe currently-available approaches for imaging single genomic loci in cells,with special focus on clus-tered regularly interspaced short palindromic repeats (CRISPR)-based imaging approaches. In addition,we discuss some of the challenges that limit the application of CRISPR-based genomic imaging approaches,and potential solutions to address these challenges. We anticipate that,with continued refinement of CRISPR-based imaging techniques,significant understanding can be gained to help decipher chromatin activities and their relevance to cellular physiology and pathogenesis.
基金supported by grants from the National Key R&D Program of China (Grant Nos. 2016YFA0501603 and 2016YFA0100702)the National Natural Science Foundation of China (Grant Nos. 31771583 and 81371613)+1 种基金the Beijing Natural Science Foundation (Grant No. 7162114)the 1000 Young Talent Program of China
文摘With numerous advancements in novel biochemical techniques, our knowledge of the role of RNAs in the regulation of cellular physiology and pathology has grown significantly over the past several decades. Nevertheless, detailed information regarding RNA processing, trafficking, and localization in living cells has been lacking due to technical limitations in imaging single RNA transcripts in living cells with high spatial and temporal resolution. In this review, we discuss tech- niques that have shown great promise for single RNA imaging, followed by highlights in our recent work in the development of molecular beacons （MBs）, a class of nanoscale oligonucleotide-probes, for detecting individual RNA transcripts in living cells. With further refinement of MB design and development of more sophisticated fluorescence microscopy techniques, we envision that MB-based approaches could promote new discoveries of RNA functions and activities.
文摘We assessed expression of IL-20 and its receptors in psoriasis,given the recent implication of IL-20 in epidermal hyperplasia.Psoriatic lesional(LS)skin consistently expressed more IL-20 mRNA than nonlesional(NL)skin.Immunoreactivity to IL-20 protein was greater in LS tissue and mainly localized to infiltrating CD68+/CD11c+(myeloid-derived)dermal leukocytes.Because this contrasted with earlier reports of a keratinocyte source,we assessed IL-20 mRNA expression in a variety of cells in vitro,and confirmed a myeloid-derived cellular source(monocytes).Plastic adhesion,activation of β2 integrins,and incubation with tumor necrosis factor-αstimulated expression in these cells.IL-20 receptor(IL-20R)αand IL-20RβmRNA was decreased in LS versus NL skin,which also contrasted with earlier findings.To investigate the relationship between IL-20 and disease activity,we examined psoriasis patients treated with the CD2-targeted agent alefacept.In therapeutic responders,lesional IL-20 mRNA decreased to NL levels,suggesting that CD2+leukocytes may proximally regulate IL-20.Finally,to assess IL-20 function,we used microarrays to screen IL-20-treated keratinocytes,which demonstrated upregulation of disease-related and IFN-γ-induced genes.Hence,IL-20 may influence inflammation through IFN-like effects.Together,these data indicate that IL-20 may be an important effector cytokine in psoriasis,and that its inhibition may represent a potential therapeutic target.
