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Dual-probe RNA FRET-FISH in Yeast

Authors: Gable M. Wadsworth, Rasesh Y. Parikh and Harold D. Kim, date: 06/05/2018, view: 233, Q&A: 0

mRNA Fluorescence In Situ Hybridization (FISH) is a technique commonly used to profile the distribution of transcripts in cells. When combined with the common single molecule technique Fluorescence Resonance Energy Transfer (FRET), FISH can ...

Single-probe RNA FISH in Yeast

Authors: Gable M. Wadsworth, Rasesh Y. Parikh and Harold D. Kim, date: 06/05/2018, view: 155, Q&A: 0

Quantitative profiling of mRNA expression is an important part of understanding the state of a cell. The technique of RNA Fluorescence In Situ Hybridization (FISH) involves targeting an RNA transcript with a set of 40 complementary ...

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Dual-probe RNA FRET-FISH in Yeast

Authors: Gable M. Wadsworth, Rasesh Y. Parikh and Harold D. Kim, date: 06/05/2018, view: 233, Q&A: 0

[Abstract] mRNA Fluorescence In Situ Hybridization (FISH) is a technique commonly used to profile the distribution of transcripts in cells. When combined with the common single molecule technique Fluorescence Resonance Energy Transfer (FRET), FISH can also be used to profile the co-expression of nearby sequences in the transcript to measure ...

Single-probe RNA FISH in Yeast

Authors: Gable M. Wadsworth, Rasesh Y. Parikh and Harold D. Kim, date: 06/05/2018, view: 155, Q&A: 0

[Abstract] Quantitative profiling of mRNA expression is an important part of understanding the state of a cell. The technique of RNA Fluorescence In Situ Hybridization (FISH) involves targeting an RNA transcript with a set of 40 complementary fluorescently labeled DNA oligonucleotide probes. However, there are many circumstances such as transcripts ...

Ectopic Gene Expression in Macrophages Using in vitro Transcribed mRNA

Authors: Pallavi Chandra and Jennifer A. Philips, date: 05/20/2018, view: 308, Q&A: 0

[Abstract] Macrophages are immune cells that contribute to host defense through various mechanisms including phagocytosis and antigen presentation. Their antimicrobial capacity is subverted by clinically important intracellular pathogens such as Mycobacterium tuberculosis. The study of host-pathogen interactions using these cells is therefore of ...

Nab Escaping AAV Mutants Isolated from Mouse Muscles

Authors: Zheng Chai, R. Jude Samulski and Chengwen Li, date: 05/05/2018, view: 297, Q&A: 0

[Abstract] Neutralizing antibodies (Nabs) are a major challenge in clinical trials of adeno-associated virus (AAV) vector gene therapy, because Nabs are able to inhibit AAV transduction in patients. We have successfully isolated several novel Nab-escaped AAV chimeric capsids in mice by administrating a mixture of AAV shuffled library and patient serum. These ...

Detecting the Interaction of Double-stranded RNA Binding Protein, Viral Protein and Primary miRNA Transcript by Co-immunoprecipitation in planta

Authors: Lijia Zheng, Chao Zhang, Jianguo Wu and Yi Li, date: 05/05/2018, view: 249, Q&A: 0

[Abstract] MicroRNAs (miRNAs) play important roles in plant growth, development, and response to infection by microbes. Double-stranded RNA binding protein 1 (DRB1) facilitates the processing of primary miRNA transcripts into mature miRNAs. Recently, we found that NS3 protein encoded by rice stripe virus (RSV) associates with DRB1 and promotes miRNA ...

Functional Evaluation of the Signal Peptides of Secreted Proteins

Authors: Weixiao Yin, Yufu Wang, Tao Chen, Yang Lin and Chaoxi Luo, date: 05/05/2018, view: 287, Q&A: 0

[Abstract] Here, we describe a method that can be used to evaluate the function of predicted signal peptides. This method utilizes the yeast Saccharomyces cerevisiae YTK12 strain and pSUC2 vector in which the pSUC2 vector with fused predicted signal peptide is transformed into yeast. The function of the signal peptides can be evaluated by using ...

Assessing the Efficacy of Small Molecule Inhibitors in a Mouse Model of Persistent Norovirus Infection

Authors: Jana Van Dycke, Johan Neyts and Joana Rocha-Pereira, date: 05/05/2018, view: 361, Q&A: 0

[Abstract] Human norovirus is the most common cause of acute gastroenteritis worldwide, resulting in estimated mortality of ~210,000 each year, of whom most are children under the age of five. However, norovirus can infect people of all age groups. There is a risk of prolonged infection in children, the elderly and patients who are immunocompromised. To ...

Purification of Total RNA from DSS-treated Murine Tissue via Lithium Chloride Precipitation

Authors: Emilie Viennois, Anika Tahsin and Didier Merlin, date: 05/05/2018, view: 343, Q&A: 0

[Abstract] We have developed a protocol to purify RNA from DSS (Dextran Sulfate Sodium)-treated mouse tissues. This method, which prevents downstream inhibition of q-RT-PCR observed in DSS-treated tissues, relies on successive precipitations with lithium chloride.

A Modified Low-quantity RNA-Seq Method for Microbial Community and Diversity Analysis Using Small Subunit Ribosomal RNA

Authors: Yong-Wei Yan, Ting Zhu, Bin Zou and Zhe-Xue Quan, date: 05/05/2018, view: 226, Q&A: 0

[Abstract] We propose a modified RNA-Seq method for small subunit ribosomal RNA (SSU rRNA)-based microbial community analysis that depends on the direct ligation of a 5’ adaptor to RNA before reverse-transcription. The method requires only a low-input quantity of RNA (10-100 ng) and does not require a DNA removal step. Using this method, we could obtain more ...

Mammalian Cell-derived Vesicles for the Isolation of Organelle Specific Transmembrane Proteins to Conduct Single Molecule Studies

Authors: Faruk H. Moonschi, Ashley M. Fox-Loe, Xu Fu and Chris I. Richards, date: 05/05/2018, view: 328, Q&A: 0

[Abstract] Cell-derived vesicles facilitate the isolation of transmembrane proteins in their physiological membrane maintaining their structural and functional integrity. These vesicles can be generated from different cellular organelles producing, housing, or transporting the proteins. Combined with single-molecule imaging, isolated organelle specific ...