RNA-sequencing (RNA-Seq) bottlenecks start with library preparation! Is it possible to prepare a RNA-Seq library in a simple and fast way? And how to avoid the bottlenecks of current RNA-sequencing library preparation?
Following on in our series on microRNAs (miRNA – Keys to open cell biology’s secret chamber) , in this post, we’ll be taking a look at Antagomirs (which are also called anti-miRs or blockmirs). These chemically engineered oligonucleotides prevent other molecules from binding to a desired site on an mRNA molecule. They’re used to silence endogenous microRNA.
In this third post of the miRNA-related series, let’s take a look at miRNA overexpression.
Short RNAs are very prone to enzymatic degradation. Novel chemical technologies now allow RNA stabilization to make them resistant to cellular RNAse attacks. However, this approach remains optimal for short term assays. LNA (locked nucleic acid) oligonucleotides show extended half life in cells, characterized by their ability to bind complementary nucleic acids with exceptional affinity. To be mentioned also, 2′-O-Me nucleotide-composed oligonucleotides – as they show increased stability against nucleases and RNA dimers with 2′-O-Me oligos are not recognized by RNAse H (which is a big advantage for antisense technologies). [Read more…]
As cell biologist, when working in labs I have in the past witnessed several “X-files” in a flask of cells along my way: all cells of an established cell line are supposed to have the same genetic background and to respond similarly to the same stimuli… but sometimes this is simply not what happens!
I would not risk saying that it all comes down to epigenetics, but much of this “unexpected” behaviour is governed at mRNA level independently of the genetic background. “Cell personality” is then defined at several levels: genetic background, epigenetics, mRNA modulation and protein/lipid content.
It’s a sophisticated web of positive/negative signals that governs cell fate and behavior. But how do cells cope with this complicated issue? [Read more…]
Nucleosomes are core particles involved in DNA packaging in eukaryotic cells. They affect gene expression levels by regulating access of DNA to transcription factors and DNA binding proteins. Nucleosome modeling is not static and chaotic. The link between nucleosome dynamics and gene expression modulation still remains a mystery, driving Life Scientists to design clever nucleosome-based in vitro assays such as those we’ll be reviewing here.
miRNA are implicated in a number of diseases and cancers. Despite the fact that their different roles in physiological and pathological cell processes are still under investigation, there is already more and more interest in considering them as future key diagnostic and prognostic biomarkers. Indeed, they are conveniently present in blood, and furthermore, their small size makes them relatively robust compared to messenger RNAs. They are well conserved into poor quality samples such as FFPE samples allowing the best hopes when exploring biobanks of tissues.
Accurate and convenient technologies aimed at profiling miRNA expression from such biobanks and blood opens up a promising era in biomarker discovery for personalized healthcare.
But still, we need to find an efficient profiling approach to cover the full miRnome that contains about 2000 Human miRNA… [Read more…]
Secrete-Pair™ dual luminescence assay kit analyzes the activities of Gaussia Luciferase (GLuc) and Secreted Alkaline Phosphatase (SEAP) as a dual-reporter system. This dual luminescence assay system includes all reagents needed for an optimal dual luminicent detection of GLuc and SEAP while allowing transfection normalization and side-by-side analysis using the same sample from the cell culture medium.
The CRISPR (Clustered, Regularly Interspaced, Short Palindromic Repeats)-Cas (CRISPR-associated) (CRISPR-Cas) system has become trendy as it is suitable for numerous applications such as gene knockouts, genome-engineering, to name but a few. In a recent Technical Bulletin, Ed Davis describes the mechanism of CRISPR-Cas for genome editing and how the recent experimental improvements improve CRISPR-Cas9 specificity while reducing off-target effects.
It’s official, CRISPR has officially reached “Breakout” status as a Google search term (see image from Google Trends). The Clustered, Regularly Interspaced, Short Palindromic Repeats (CRISPR) and RNA-guided Cas9 nucleases are an exciting new gene editing tool that allows life science researchers to directly modify the DNA of the organism they study.