scholarly journals Reverse genetics demonstrate the role of mucosal C-type lectins in food particle selection in the oysterCrassostrea virginica

2018 ◽  
Vol 221 (6) ◽  
pp. jeb174094 ◽  
Author(s):  
Emmanuelle Pales Espinosa ◽  
Bassem Allam
Keyword(s):  
Reverse Genetics ◽  
Food Particle ◽  
Particle Selection

scholarly journals Centrin2 regulates CP110 removal in primary cilium formation

2015 ◽  
Vol 208 (6) ◽  
pp. 693-701 ◽  
Author(s):  
Suzanna L. Prosser ◽  
Ciaran G. Morrison
Keyword(s):  
Calcium Binding ◽  
Primary Cilia ◽  
Reverse Genetics ◽  
Cell Types ◽  
Serum Starvation ◽  
Basal Bodies ◽  
Cilium Formation ◽  
Cellular Quiescence ◽  
Retinal Pigmented Epithelial Cells

Primary cilia are antenna-like sensory microtubule structures that extend from basal bodies, plasma membrane–docked mother centrioles. Cellular quiescence potentiates ciliogenesis, but the regulation of basal body formation is not fully understood. We used reverse genetics to test the role of the small calcium-binding protein, centrin2, in ciliogenesis. Primary cilia arise in most cell types but have not been described in lymphocytes. We show here that serum starvation of transformed, cultured B and T cells caused primary ciliogenesis. Efficient ciliogenesis in chicken DT40 B lymphocytes required centrin2. We disrupted CETN2 in human retinal pigmented epithelial cells, and despite having intact centrioles, they were unable to make cilia upon serum starvation, showing abnormal localization of distal appendage proteins and failing to remove the ciliation inhibitor CP110. Knockdown of CP110 rescued ciliation in CETN2-deficient cells. Thus, centrin2 regulates primary ciliogenesis through controlling CP110 levels.

scholarly journals The Hemagglutinin-Neuraminidase Protein of Newcastle Disease Virus Determines Tropism and Virulence

2004 ◽  
Vol 78 (8) ◽  
pp. 4176-4184 ◽  
Author(s):  
Zhuhui Huang ◽  
Aruna Panda ◽  
Subbiah Elankumaran ◽  
Dhanasekaran Govindarajan ◽  
Daniel D. Rockemann ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Reverse Genetics ◽  
Index Test ◽  
Death Time ◽  
Virulent Virus ◽  
Hn Gene ◽  
Chimeric Viruses

ABSTRACT The hemagglutinin-neuraminidase (HN) protein of Newcastle disease virus (NDV) plays a crucial role in the process of infection. However, the exact contribution of the HN gene to NDV pathogenesis is not known. In this study, the role of the HN gene in NDV virulence was examined. By use of reverse genetics procedures, the HN genes of a virulent recombinant NDV strain, rBeaudette C (rBC), and an avirulent recombinant NDV strain, rLaSota, were exchanged. The hemadsorption and neuraminidase activities of the chimeric viruses showed significant differences from those of their parental strains, but heterotypic F and HN pairs were equally effective in fusion promotion. The tissue tropism of the viruses was shown to be dependent on the origin of the HN protein. The chimeric virus with the HN protein derived from the virulent virus exhibited a tissue predilection similar to that of the virulent virus, and vice versa. The chimeric viruses with reciprocal HN proteins either gained or lost virulence, as determined by a standard intracerebral pathogenicity index test of chickens and by the mean death time in chicken embryos (a measure devised to classify these viruses), indicating that virulence is a function of the amino acid differences in the HN protein. These results are consistent with the hypothesis that the virulence of NDV is multigenic and that the cleavability of F protein alone does not determine the virulence of a strain.

Role of rheological characteristics in amorphous food particle-wall collisions in spray drying

2010 ◽  
Vol 198 (2) ◽  
pp. 251-257 ◽  
Author(s):  
M.W. Woo ◽  
W.R.W. Daud ◽  
A.S. Mujumdar ◽  
S.M. Tasirin ◽  
M.Z.M. Talib
Keyword(s):  
Spray Drying ◽  
Rheological Characteristics ◽  
Food Particle

Anthelmintic metabolism in parasitic helminths: proteomic insights

Parasitology ◽  
2012 ◽  
Vol 139 (9) ◽  
pp. 1205-1217 ◽  
Author(s):  
PETER M. BROPHY ◽  
NEIL MACKINTOSH ◽  
RUSSELL M. MORPHEW
Keyword(s):  
Reverse Genetics ◽  
Anthelmintic Resistance ◽  
Phase Iii ◽  
Study Phase ◽  
Post Translational Modification ◽  
Cellular Mechanisms ◽  
Parasitic Helminths ◽  
Global Threat ◽  
Cytochrome P 450

SUMMARYAnthelmintics are the cornerstone of parasitic helminth control. Surprisingly, understanding of the biochemical pathways used by parasitic helminths to detoxify anthelmintics is fragmented, despite the increasing global threat of anthelmintic resistance within the ruminant and equine industries. Reductionist biochemistry has likely over-estimated the enzymatic role of glutathione transferases in anthelmintic metabolism and neglected the potential role of the cytochrome P-450 superfamily (CYPs). Proteomic technologies offers the opportunity to support genomics, reverse genetics and pharmacokinetics, and provide an integrated insight into both the cellular mechanisms underpinning response to anthelmintics and also the identification of biomarker panels for monitoring the development of anthelmintic resistance. To date, there have been limited attempts to include proteomics in anthelmintic metabolism studies. Optimisations of membrane, post-translational modification and interaction proteomic technologies in helminths are needed to especially study Phase I CYPs and Phase III ABC transporter pumps for anthelmintics and their metabolites.

scholarly journals Breathing with Phox2b

2009 ◽  
Vol 364 (1529) ◽  
pp. 2477-2483 ◽  
Author(s):  
Véronique Dubreuil ◽  
Jacques Barhanin ◽  
Christo Goridis ◽  
Jean-François Brunet
Keyword(s):  
Reverse Genetics ◽  
Human Genetics ◽  
Respiratory Rhythm ◽  
Neuronal Population ◽  
Cellular Level ◽  
Small Population ◽  
Respiratory Neurons ◽  
Retrotrapezoid Nucleus ◽  
Parafacial Respiratory Group

In the last few years, elucidation of the architecture of breathing control centres has reached the cellular level. This has been facilitated by increasing knowledge of the molecular signatures of various classes of hindbrain neurons. Here, we review the advances achieved by studying the homeodomain factor Phox2b , a transcriptional determinant of neuronal identity in the central and peripheral nervous systems. Evidence from human genetics, neurophysiology and mouse reverse genetics converges to implicate a small population of Phox2b -dependent neurons, located in the retrotrapezoid nucleus, in the detection of CO 2 , which is a paramount source of the ‘drive to breathe’. Moreover, the same and other studies suggest that an overlapping or identical neuronal population, the parafacial respiratory group, might contribute to the respiratory rhythm at least in some circumstances, such as for the initiation of breathing following birth. Together with the previously established Phox2b dependency of other respiratory neurons (which we review briefly here), our new data highlight a key role of this transcription factor in setting up the circuits for breathing automaticity.

scholarly journals Identification of maternal-effect genes in zebrafish using maternal crispants

Development ◽  
2021 ◽  
Author(s):  
Cara E. Moravec ◽  
Gabriella C. Voit ◽  
Jarred Otterlee ◽  
Francisco Pelegri
Keyword(s):  
Maternal Effect ◽  
Reverse Genetics ◽  
Germ Line ◽  
Maternal Factors ◽  
Single Generation ◽  
Maternal Effect Genes ◽  
Biallelic Mutations ◽  
Genome Activation

In animals, early development is dependent on a pool of maternal factors, both RNA and proteins, which are required for basic cellular process and cell differentiation until zygotic genome activation. The role of a majority of these maternally expressed factors is not fully understood. By exploiting the biallelic editing ability of CRISPR-Cas9, we identify and characterize maternal-effect genes in a single generation, using a maternal crispant technique. We validated the ability to generate biallelic mutations in the germline by creating maternal crispants that phenocopied previously characterized maternal-effect genes: motley/birc5b, tmi/prc1l, and aura/mid1ip1. Additionally, by targeting maternally expressed genes of unknown function in zebrafish, we identified two new maternal-effect zebrafish genes, kpna7 and fhdc3. The genetic identity of these maternal crispants was confirmed by sequencing haploid progeny from F0 females, which allowed the analysis of newly induced lesions in the maternal germ line. Our studies show that maternal crispants allow for the effective identification and primary characterization of maternal-effect genes in a single generation, facilitating the reverse genetics analysis of maternal factors that drive embryonic development.

scholarly journals N-glycosylation of infectious bronchitis virus M41 spike determines receptor specificity

2020 ◽  
Vol 101 (6) ◽  
pp. 599-608
Author(s):  
K. M. Bouwman ◽  
N. Habraeken ◽  
A. Laconi ◽  
A. J. Berends ◽  
L. Groenewoud ◽  
...  
Keyword(s):  
Infectious Bronchitis Virus ◽  
Reverse Genetics ◽  
Glycosylation Site ◽  
Host Cells ◽  
Receptor Specificity ◽  
Knock Out ◽  
Recombinant Viruses ◽  
Infectious Bronchitis

Infection of chicken coronavirus infectious bronchitis virus (IBV) is initiated by binding of the viral heavily N-glycosylated attachment protein spike to the alpha-2,3-linked sialic acid receptor Neu5Ac. Previously, we have shown that N-glycosylation of recombinantly expressed receptor binding domain (RBD) of the spike of IBV-M41 is of critical importance for binding to chicken trachea tissue. Here we investigated the role of N-glycosylation of the RBD on receptor specificity and virus replication in the context of the virus particle. Using our reverse genetics system we were able to generate recombinant IBVs for nine-out-of-ten individual N-glycosylation mutants. In vitro growth kinetics of these viruses were comparable to the virus containing the wild-type M41-S1. Furthermore, Neu5Ac binding by the recombinant viruses containing single N-glycosylation site knock-out mutations matched the Neu5Ac binding observed with the recombinant RBDs. Five N-glycosylation mutants lost the ability to bind Neu5Ac and gained binding to a different, yet unknown, sialylated glycan receptor on host cells. These results demonstrate that N-glycosylation of IBV is a determinant for receptor specificity.

Sign in / Sign up

Export Citation Format

Share Document