There is a growing realization that cell-to-cell variations in gene expression have importantbiological consequences underlying phenotype diversity and cell fate. Although analytical toolsfor measuring gene expression...There is a growing realization that cell-to-cell variations in gene expression have importantbiological consequences underlying phenotype diversity and cell fate. Although analytical toolsfor measuring gene expression, such as DNA microarrays, reverse-transcriptase PCR and in situhybridization have been widely utilized to discover the role of genetic variations in governingcellular behavior, these methods are performed in cell lysates and/or on fixed cells, and thereforelack the ability to provide comprehensive spatial-dynamic information on gene expression. Thishas invoked the recent development of molecular imaging strategies capable of illuminatingthe distribution and dynamics of RNA molecules in living cells. In this review, we describe aclass of molecular imaging probes known as molecular beacons (MBs), which have increasinglybecome the probe of choice for imaging RNA in living cells. In addition, we present the majorchallenges that can limit the ability of MBs to provide accurate measurements of RNA, anddiscuss efforts that have been made to overcome these challenges. It is envisioned that withcontinued refinement of the MB design, MBs will eventually become an indispensable tool foranalyzing gene expression in biology and medicine.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
基金the National Institutes of Health(NCI)R21 CA116102 and R21 CA125088the National Science Foundation BES-0616031,and the American Cancer Society RSG-07-005-01.
文摘There is a growing realization that cell-to-cell variations in gene expression have importantbiological consequences underlying phenotype diversity and cell fate. Although analytical toolsfor measuring gene expression, such as DNA microarrays, reverse-transcriptase PCR and in situhybridization have been widely utilized to discover the role of genetic variations in governingcellular behavior, these methods are performed in cell lysates and/or on fixed cells, and thereforelack the ability to provide comprehensive spatial-dynamic information on gene expression. Thishas invoked the recent development of molecular imaging strategies capable of illuminatingthe distribution and dynamics of RNA molecules in living cells. In this review, we describe aclass of molecular imaging probes known as molecular beacons (MBs), which have increasinglybecome the probe of choice for imaging RNA in living cells. In addition, we present the majorchallenges that can limit the ability of MBs to provide accurate measurements of RNA, anddiscuss efforts that have been made to overcome these challenges. It is envisioned that withcontinued refinement of the MB design, MBs will eventually become an indispensable tool foranalyzing gene expression in biology and medicine.
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.
