Biology subjects

Wang, J. W.

Publications and source records attributed to Wang, J. W..

5 recordsLinked to original sources

bioRxiv2021

Polymer-Conjugated Carbon Nanotubes for Biomolecule Loading

Nanomaterials have emerged as an invaluable tool for the delivery of biomolecules such as DNA and RNA, with various applications in genetic engineering and post-transcriptional genetic manipulation. Alongside this development, there has been an increasing use of polymer-based techniques, such as polyethyleneimine (PEI), to electrostatically load polynucleotide cargoes onto nanomaterial carriers. However, there remains a need to assess nanomaterial properties, conjugation conditions, and biocompatibility of these nanomaterial-polymer constructs, particularly for use in plant systems. In this work, we develop mechanisms to optimize DNA loading on single-walled carbon nanotubes (SWNTs) with a library of polymer-SWNT constructs and assess DNA loading ability, polydispersity, and both chemical and colloidal stability. Counterintuitively, we demonstrate that polymer hydrolysis from nanomaterial surfaces can occur depending on polymer properties and attachment chemistries, and describe mitigation strategies against construct degradation. Given the growing interest in delivery applications in plant systems, we also assess the toxicity of polymer-based nanomaterials in plants and provide recommendations for future design of nanomaterial-based polynucleotide delivery strategies.

biochemistry↗

bioRxiv2021

Supervised Learning Model Predicts Protein Adsorption to Nanotubes

Engineered nanoparticles are advantageous for numerous biotechnology applications, including biomolecular sensing and delivery. However, testing the compatibility and function of nanotechnologies in biological systems requires a heuristic approach, where unpredictable biofouling via protein corona formation often prevents effective implementation. Moreover, rational design of biomolecule-nanoparticle conjugates requires prior knowledge of such interactions or extensive experimental testing. Toward better applying engineered nanoparticles in biological systems, herein, we develop a random forest classifier (RFC) trained with proteomic mass spectrometry data that identifies proteins that adsorb to nanoparticles, based solely on the proteins amino acid sequence. We model proteins that populate the corona of a single-walled carbon nanotube (SWCNT)-based optical nanosensor and study whether there is a relationship between the proteins amino acid-based properties and the proteins adsorption to SWCNTs. We optimize the classifier and characterize the classifier performance against other models. To evaluate the predictive power of our model, we apply the classifier to rapidly identify proteins with high binding affinity to SWCNTs, followed by experimental validation. We further determine protein features associated with increased likelihood of SWCNT binding: high content of solvent-exposed glycine residues and non-secondary structure-associated amino acids. Conversely, proteins with high content of leucine residues and beta-sheet-associated amino acids are less likely to form the SWCNT protein corona. The classifier presented herein provides a step toward undertaking the otherwise intractable problem of predicting protein-nanoparticle interactions, which is needed for more rapid and effective translation of nanobiotechnologies from in vitro synthesis to in vivo use. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=66 SRC="FIGDIR/small/449132v2_ufig1.gif" ALT="Figure 1"> View larger version (22K): org.highwire.dtl.DTLVardef@bbc175org.highwire.dtl.DTLVardef@9a0b94org.highwire.dtl.DTLVardef@16e22e1org.highwire.dtl.DTLVardef@1b40a84_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗

bioRxiv2020

Groucho/TLE opposes axial to hypaxial motor neuron development

Spinal cord motor neuron diversity and the ensuing variety of motor circuits allow for the processing of elaborate muscular behaviours such as body posture and breathing. Little is known, however, about the molecular mechanisms behind the specification of axial and hypaxial motor neurons controlling postural and respiratory functions respectively. Here we show that the Groucho/TLE (TLE) transcriptional corepressor is a multi-step regulator of axial and hypaxial motor neuron diversification in the developing spinal cord. TLE first promotes axial motor neuron specification at the expense of hypaxial identity by cooperating with non-canonical WNT5A signalling within the motor neuron progenitor domain. TLE further acts during post-mitotic motor neuron diversification to promote axial motor neuron topology and axonal connectivity whilst suppressing hypaxial traits. These findings provide evidence for essential and sequential roles of TLE in the spatial and temporal coordination of events regulating the development of motor neurons influencing posture and controlling respiration. HIGHLIGHTSO_LIGroucho/TLE mediates non-canonical WNT signalling in developing motor neurons C_LIO_LINon canonical WNT:TLE pathway regulates thoracic motor neuron diversification C_LIO_LITLE promotes axial while inhibiting hypaxial motor neuron development C_LIO_LITLE influences developing motor neuron topology and muscle innervation C_LI IN BRIEFSalin-Cantegrel et al use in ovo engineered approaches to show that a non-canonical WNT:TLE pathway coordinates temporally and spatially separated elements of motor neuron diversification, repressing hypaxial motor neuron development to promote the axial fate. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=116 SRC="FIGDIR/small/986323v1_ufig1.gif" ALT="Figure 1"> View larger version (28K): org.highwire.dtl.DTLVardef@e2d96dorg.highwire.dtl.DTLVardef@130baf4org.highwire.dtl.DTLVardef@41fa32org.highwire.dtl.DTLVardef@190ebd3_HPS_FORMAT_FIGEXP M_FIG TLE contribution to the development of thoracic somatic motor columns Progenitor cells in the ventral pMN domain are exposed to higher concentrations of non-canonical WNTs and express more TLE. Cooperation of non-canonical WNTs and TLE renders ventral pMN progenitors refractory to a respiratory MN fate, thereby contributing to the separation of MMC and RMC MN lineages. Differentiating MNs that maintain high TLE expression also maintain LHX3 expression, adopt axial motor neuron topology and connect to axial muscles. TLE activity in differentiating MMC MNs prevents the acquisition of respiratory MN topology and innervation traits. C_FIG

neuroscience↗

bioRxiv2018

Sphingosine-1-Phosphate (S1P) analogue Phyto-sphingosine-1-Phosphate (P1P) improves the in vitro maturation efficiency of porcine oocytes via regulation of oxidative stress and apoptosis

Phytosphingosine-1-Phosphate (P1P) is a signaling sphingolipid regulating various physiological activities. Yet, little is known of the effect of P1P in the context of reproduction. As such, we aimed to investigate the influence of P1P on oocyte maturation during porcine in vitro maturation (IVM). Here we report the expression of S1PR1-3 among P1P receptors (S1PR1-4) in cumulus cells and oocytes. When P1P was treated by concentrations 10 nM, 50 nM, 100 nM, and 1000 nM during IVM, Metaphase II rate was significantly increased in 1000 nM (=1 M) P1P treatment group. Maturation rate improvement by P1P supplementation was only observed in the presence of EGF. Oocytes under the influence of P1P decreased intracellular ROS levels yet did not show significant differences in GSH levels. In our molecular studies, P1P treatment up-regulated gene expressions involved in cumulus expansion (Has2 and EGF), antioxidant enzyme (SOD3 and Cat), and developmental competence (Oct4) while activating ERK1/2 and Akt signaling. P1P treatment also influenced oocyte survival by shifting the ratio of Bcl-2 to Bax, while inactivating JNK signaling. We further demonstrated that oocytes matured with P1P significantly displayed not only higher developmental competence (cleavage and blastocyst formation rate), but also greater blastocyst quality (total cell number and the ratio of apoptotic cells) when activated via parthenogenetic activation (PA) and in vitro fertilization (IVF). Despite low levels of endogenous P1P found in animals, exogenous P1P was able to influence animal reproduction as shown by increased porcine oocyte maturation as well as preimplantation embryo development.

developmental biology↗

bioRxiv2017

A Versatile Genetic Tool For Post-Translational Control Of Gene Expression With A Small Molecule In Drosophila melanogaster

Several techniques have been developed in Drosophila to control gene expression temporally. While some of these techniques are incompatible with existing GAL4 lines, others suffer from side effects on physiology or behavior. Here, we describe a method of post-translational temporal control of gene expression which is compatible with the current library of transgenic reagents. We adopted a strategy to regulate protein degradation by fusing a protein of interest to a destabilizing domain (DD) derived from the Escherichia coli dihydrofolate reductase (ecDHFR). Trimethoprim (TMP), a stabilizing small molecule, binds to DD and blocks degradation of the chimeric protein. With a GFP-DD reporter, we show that this system is effective across different tissues and developmental stages in the fly. Notably, feeding flies with TMP can increase the expression level of GFP-DD up to 34 times in a dosage-dependent and reversible manner without altering the lifespan or behavior of the animal. To broaden the utility of our method, we engineered GAL80-DD flies that can be crossed to the available GAL4 lines to control the temporal pattern of gene expression with TMP. We also developed an inducible recombinase, FLP-DD, for high-efficiency sparse labeling and intersectional lineage analysis. Finally, we demonstrated the utility of the DD system in manipulating neuronal activity of sensory neurons. In summary, we have developed a system to control in vivo gene expression levels with negligible background, large dynamic range, and in a reversible manner, all by feeding a small molecule to Drosophila melanogaster.

genetics↗