Human Genetics

Author(s):  
Antonio Regalado

Genetic research is moving faster than a nematode poked by a platinum needle. Every week, the scientific journals report a score of new gene discoveries made in mice, worms, and men. How can a science journalist cover it all? It's hopeless, of course. So one thing I always keep in mind is it's often the methods or scientific tools behind these molecular discoveries, not the discoveries themselves, that present the best story possibilities. Examples of topics for such “tool stories” include DNA chips, proteomics, and new imaging technologies like the green-fluorescent protein used to make zebrafish and other laboratory critters glow. In writing about the technologies that drive biological research, I've found a formula that has worked well for me, time and again. Of course, not every story fits the same mold, and the best ones break it. But it's important to be familiar with how a tool story typically comes to be, and how to write one. I like to think about biology as a big onion that's rapidly being peeled. There are tens of thousands of biologists peeling away every day, figuring out all of life's working parts. But I never saw much sense in inspecting every peel for its news potential. (And some editors I know refer dismissively to the latest uncovering of a gene for heart attack or schizophrenia as “gene-of-the-week” stories.) It's better, sometimes, to focus on the new techniques and ideas for peeling the onion. Tool stories are big-picture stories that can be newsy, but the trends tend to have a long shelf life. They endure through numerous news cycles, and ultimately nearly every outlet in the journalistic food chain will cover the big ones. Your decision is when to catch the wave. Some reporters put a big emphasis on being first, but others will be content to watch the story unfold and cover their piece of it when it's right for whatever market they happen to be writing for. Either way, a tale of how a new technology is changing biological research is a great way to teach your readers—and yourself—about how science really works.

2021 ◽  
Vol 7 (2) ◽  
pp. eabd2529
Author(s):  
Kazuki Okamoto ◽  
Teppei Ebina ◽  
Naoki Fujii ◽  
Kuniaki Konishi ◽  
Yu Sato ◽  
...  

Optical investigation and manipulation constitute the core of biological experiments. Here, we introduce a new borosilicate glass material that contains the rare-earth ion terbium(III) (Tb3+), which emits green fluorescence upon blue light excitation, similar to green fluorescent protein (GFP), and thus is widely compatible with conventional biological research environments. Micropipettes made of Tb3+-doped glass allowed us to target GFP-labeled cells for single-cell electroporation, single-cell transcriptome analysis (Patch-seq), and patch-clamp recording under real-time fluorescence microscopic control. The glass also exhibited potent third harmonic generation upon infrared laser excitation and was usable for online optical targeting of fluorescently labeled neurons in the in vivo neocortex. Thus, Tb3+-doped glass simplifies many procedures in biological experiments.


1999 ◽  
Vol 10 (12) ◽  
pp. 4311-4326 ◽  
Author(s):  
Barth Grant ◽  
David Hirsh

The Caenorhabditis elegans oocyte is a highly amenable system for forward and reverse genetic analysis of receptor-mediated endocytosis. We describe the use of transgenic strains expressing a vitellogenin::green fluorescent protein (YP170::GFP) fusion to monitor yolk endocytosis by theC. elegans oocyte in vivo. This YP170::GFP reporter was used to assay the functions of C. eleganspredicted proteins homologous to vertebrate endocytosis factors using RNA-mediated interference. We show that the basic components and pathways of endocytic trafficking are conserved between C. elegans and vertebrates, and that this system can be used to test the endocytic functions of any new gene. We also used the YP170::GFP assay to identify rme(receptor-mediated endocytosis) mutants. We describe a new member of the low-density lipoprotein receptor superfamily, RME-2, identified in our screens for endocytosis defective mutants. We show that RME-2 is the C. elegans yolk receptor.


2010 ◽  
Vol 9 (5) ◽  
pp. 827-830 ◽  
Author(s):  
Julian Lai ◽  
Seng Kah Ng ◽  
Fang Fang Liu ◽  
Rajesh Narhari Patkar ◽  
Yanfen Lu ◽  
...  

ABSTRACT A new gene-tagging method (marker fusion tagging [MFT]) is demonstrated for Neurospora crassa and Magnaporthe oryzae. Translational fusions between the hygromycin B resistance gene and various markers are inserted into genes of interest by homologous recombination to produce chromosomally encoded fusion proteins. This method can produce tags at any position and create deletion alleles that maintain N- and C-terminal sequences. We show the utility of MFT by producing enhanced green fluorescent protein (EGFP) tags in proteins localized to nuclei, spindle pole bodies, septal pore plugs, Woronin bodies, developing septa, and the endoplasmic reticulum.


2008 ◽  
Vol 2008 ◽  
pp. 1-12 ◽  
Author(s):  
Mee-Jung Han ◽  
Jeong Wook Lee ◽  
Sang Yup Lee ◽  
Jong Shin Yoo

InEscherichia coli, a long-chain acyl-CoA is a regulatory signal that modulates gene expression through its binding to a transcription factor FadR. In this study, comparative proteomic analysis ofE. coliin the presence of glucose and oleic acid was performed to understand cell physiology in response to oleic acid. Among total of 52 proteins showing altered expression levels with oleic acid presence, 9 proteins including AldA, Cdd, FadA, FadB, FadL, MalE, RbsB, Udp, and YccU were newly synthesized. Among the genes that were induced by oleic acid, the promoter of thealdAgene was used for the production of a green fluorescent protein (GFP). Analysis of fluorescence intensities and confocal microscopic images revealed that soluble GFP was highly expressed under the control of thealdApromoter. These results suggest that proteomics is playing an important role not only in biological research but also in various biotechnological applications.


2019 ◽  
Author(s):  
Chi-Yun Lin ◽  
Matthew Romei ◽  
Luke Oltrogge ◽  
Irimpan Mathews ◽  
Steven Boxer

Green fluorescent protein (GFPs) have become indispensable imaging and optogenetic tools. Their absorption and emission properties can be optimized for specific applications. Currently, no unified framework exists to comprehensively describe these photophysical properties, namely the absorption maxima, emission maxima, Stokes shifts, vibronic progressions, extinction coefficients, Stark tuning rates, and spontaneous emission rates, especially one that includes the effects of the protein environment. In this work, we study the correlations among these properties from systematically tuned GFP environmental mutants and chromophore variants. Correlation plots reveal monotonic trends, suggesting all these properties are governed by one underlying factor dependent on the chromophore's environment. By treating the anionic GFP chromophore as a mixed-valence compound existing as a superposition of two resonance forms, we argue that this underlying factor is defined as the difference in energy between the two forms, or the driving force, which is tuned by the environment. We then introduce a Marcus-Hush model with the bond length alternation vibrational mode, treating the GFP absorption band as an intervalence charge transfer band. This model explains all the observed strong correlations among photophysical properties; related subtopics are extensively discussed in Supporting Information. Finally, we demonstrate the model's predictive power by utilizing the additivity of the driving force. The model described here elucidates the role of the protein environment in modulating photophysical properties of the chromophore, providing insights and limitations for designing new GFPs with desired phenotypes. We argue this model should also be generally applicable to both biological and non-biological polymethine dyes.<br>


2019 ◽  
Author(s):  
Jeffrey Chang ◽  
Matthew Romei ◽  
Steven Boxer

<p>Double-bond photoisomerization in molecules such as the green fluorescent protein (GFP) chromophore can occur either via a volume-demanding one-bond-flip pathway or via a volume-conserving hula-twist pathway. Understanding the factors that determine the pathway of photoisomerization would inform the rational design of photoswitchable GFPs as improved tools for super-resolution microscopy. In this communication, we reveal the photoisomerization pathway of a photoswitchable GFP, rsEGFP2, by solving crystal structures of <i>cis</i> and <i>trans</i> rsEGFP2 containing a monochlorinated chromophore. The position of the chlorine substituent in the <i>trans</i> state breaks the symmetry of the phenolate ring of the chromophore and allows us to distinguish the two pathways. Surprisingly, we find that the pathway depends on the arrangement of protein monomers within the crystal lattice: in a looser packing, the one-bond-flip occurs, whereas in a tighter packing (7% smaller unit cell size), the hula-twist occurs.</p><p> </p><p> </p><p> </p><p> </p><p> </p><p> </p> <p> </p>


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