scholarly journals A promising cell line for research on the control of locust plagues

2021 ◽  
Author(s):  
Kazuyo Watanabe ◽  
Ryohei Sugahara ◽  
Seiji Tanaka ◽  
Takumi Kayukawa
Keyword(s):  
Cell Line ◽  
High Throughput Screening ◽  
Large Scale ◽  
Target Genes ◽  
Schistocerca Gregaria ◽  
Molecular Biological Analysis ◽  
Low Concentrations ◽  
Rnai Efficiency ◽  
Candidate Target ◽  
A Cell

Abstract The desert locust (Schistocerca gregaria), which forms a huge swarm, consuming large amounts of wild and agricultural plants, is the most destructive migratory pest in the world. Recently, a large-scale locust plague occurred from Africa to South Asia. The main methods used to control these pests, which involve the use of inexpensive and highly residual insecticides, are raising concerns on their impact on humans, domestic animals, and the environment. Cell lines are a useful research tool in molecular biological analysis and drug discovery; however, no cell line is yet available for the research of S. gregaria. Here, we succeeded in establishing a cell line (Sg-155 cells) from S. gregaria embryos and validated its utility. Soaking with low concentrations of dsRNA induced high and long-lasting RNAi efficiency in Sg-155 cells. Furthermore, response to an insect hormone, a candidate target as an anti-locust agent, was observed at the gene expression level. Thus, the Sg-155 cell line is useful in exploring and evaluating target genes and could therefore be applied as a high-throughput screening tool in the development of anti-locust agents.

scholarly journals Identification of a juvenile-hormone signaling inhibitor via high-throughput screening of a chemical library

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Takumi Kayukawa ◽  
Kenjiro Furuta ◽  
Keisuke Nagamine ◽  
Tetsuro Shinoda ◽  
Kiyoaki Yonesu ◽  
...  
Keyword(s):  
Juvenile Hormone ◽  
Cell Line ◽  
Signaling Pathway ◽  
High Throughput ◽  
High Throughput Screening ◽  
Large Scale ◽  
Ex Vivo ◽  
Agricultural Field ◽  
Chemical Library ◽  
Field Development

Abstract Insecticide resistance has recently become a serious problem in the agricultural field. Development of insecticides with new mechanisms of action is essential to overcome this limitation. Juvenile hormone (JH) is an insect-specific hormone that plays key roles in maintaining the larval stage of insects. Hence, JH signaling pathway is considered a suitable target in the development of novel insecticides; however, only a few JH signaling inhibitors (JHSIs) have been reported, and no practical JHSIs have been developed. Here, we established a high-throughput screening (HTS) system for exploration of novel JHSIs using a Bombyx mori cell line (BmN_JF&AR cells) and carried out a large-scale screening in this cell line using a chemical library. The four-step HTS yielded 69 compounds as candidate JHSIs. Topical application of JHSI48 to B. mori larvae caused precocious metamorphosis. In ex vivo culture of the epidermis, JHSI48 suppressed the expression of the Krüppel homolog 1 gene, which is directly activated by JH-liganded receptor. Moreover, JHSI48 caused a parallel rightward shift in the JH response curve, suggesting that JHSI48 possesses a competitive antagonist-like activity. Thus, large-scale HTS using chemical libraries may have applications in development of future insecticides targeting the JH signaling pathway.

scholarly journals Meis2 Is Required for Inner Ear Formation and Proper Morphogenesis of the Cochlea

2021 ◽  
Vol 9 ◽  
Author(s):  
María Beatriz Durán Alonso ◽  
Victor Vendrell ◽  
Iris López-Hernández ◽  
María Teresa Alonso ◽  
Donna M. Martin ◽  
...  
Keyword(s):  
Nervous System ◽  
Inner Ear ◽  
Cell Line ◽  
Peripheral Nervous System ◽  
Target Genes ◽  
Otic Vesicle ◽  
Vesicle Formation ◽  
Tissue Specific ◽  
Cochlear Duct ◽  
Candidate Target

Meis genes have been shown to control essential processes during development of the central and peripheral nervous system. Here we have explored the roles of the Meis2 gene during vertebrate inner ear induction and the formation of the cochlea. Meis2 is expressed in several tissues required for inner ear induction and in non-sensory tissue of the cochlear duct. Global inactivation of Meis2 in the mouse leads to a severely reduced size of the otic vesicle. Tissue-specific knock outs of Meis2 reveal that its expression in the hindbrain is essential for otic vesicle formation. Inactivation of Meis2 in the inner ear itself leads to an aberrant coiling of the cochlear duct. By analyzing transcriptomes obtained from Meis2 mutants and ChIPseq analysis of an otic cell line, we define candidate target genes for Meis2 which may be directly or indirectly involved in cochlear morphogenesis. Taken together, these data show that Meis2 is essential for inner ear formation and provide an entry point to unveil the network underlying proper coiling of the cochlear duct.

scholarly journals Identification of Potential Pharmacoperones Capable of Rescuing the Functionality of Misfolded Vasopressin 2 Receptor Involved in Nephrogenic Diabetes Insipidus

2016 ◽  
Vol 21 (8) ◽  
pp. 824-831 ◽  
Author(s):  
Emery Smith ◽  
Jo Ann Janovick ◽  
Thomas D. Bannister ◽  
Justin Shumate ◽  
Louis Scampavia ◽  
...  
Keyword(s):  
Diabetes Insipidus ◽  
High Throughput ◽  
High Throughput Screening ◽  
Nephrogenic Diabetes Insipidus ◽  
Large Scale ◽  
Detection System ◽  
System A ◽  
Protein Mutants ◽  
A Cell ◽  
Homogeneous Cell

Pharmacoperones correct the folding of otherwise misfolded protein mutants, restoring function (i.e., providing “rescue”) by correcting their trafficking. Currently, most pharmacoperones possess intrinsic antagonist activity because they were identified using methods initially aimed at discovering such functions. Here, we describe an ultra-high-throughput homogeneous cell-based assay with a cAMP detection system, a method specifically designed to identify pharmacoperones of the vasopressin type 2 receptor (V2R), a GPCR that, when mutated, is associated with nephrogenic diabetes insipidus. Previously developed methods to identify compounds capable of altering cellular trafficking of V2R were modified and used to screen a 645,000 compound collection by measuring the ability of library compounds to rescue a mutant hV2R [L83Q], using a cell-based luminescent detection system. The campaign initially identified 3734 positive modulators of cAMP. The confirmation and counterscreen identified only 147 of the active compounds with an EC50 of ≤5 µM. Of these, 83 were reconfirmed as active through independently obtained pure samples and were also inactive in a relevant counterscreen. Active and tractable compounds within this set can be categorized into three predominant structural clusters, described here, in the first report detailing the results of a large-scale pharmacoperone high-throughput screening campaign.

scholarly journals Development of a Cell-Based High-Throughput Screening Assay to Identify Porcine Host Defense Peptide-Inducing Compounds

2018 ◽  
Vol 2018 ◽  
pp. 1-13 ◽  
Author(s):  
Zhuo Deng ◽  
Jing Wang ◽  
Wentao Lyu ◽  
Xuwen Wieneke ◽  
Robert Matts ◽  
...  
Keyword(s):  
Cell Line ◽  
High Throughput ◽  
High Throughput Screening ◽  
Hdac Inhibitors ◽  
Luciferase Reporter ◽  
Reporter Cell Line ◽  
Reporter Cell ◽  
Screening Assay ◽  
High Throughput Screening Assay ◽  
A Cell

Novel alternatives to antibiotics are needed for the swine industry, given increasing restrictions on subtherapeutic use of antibiotics. Augmenting the synthesis of endogenous host defense peptides (HDPs) has emerged as a promising antibiotic-alternative approach to disease control and prevention. To facilitate the identification of HDP inducers for swine use, we developed a stable luciferase reporter cell line, IPEC-J2/PBD3-luc, through permanent integration of a luciferase reporter gene driven by a 1.1 kb porcine β-defensin 3 (PBD3) gene promoter in porcine IPEC-J2 intestinal epithelial cells. Such a stable reporter cell line was employed in a high-throughput screening of 148 epigenetic compounds and 584 natural products, resulting in the identification of 41 unique hits with a minimum strictly standardized mean difference (SSMD) value of 3.0. Among them, 13 compounds were further confirmed to give at least a 5-fold increase in the luciferase activity in the stable reporter cell line, with 12 being histone deacetylase (HDAC) inhibitors. Eight compounds were subsequently observed to be comparable to sodium butyrate in inducing PBD3 mRNA expression in parental IPEC-J2 cells in the low micromolar range. Six HDAC inhibitors including suberoylanilide hydroxamine (SAHA), HC toxin, apicidin, panobinostat, SB939, and LAQ824 were additionally found to be highly effective HDP inducers in a porcine 3D4/31 macrophage cell line. Besides PBD3, other HDP genes such as PBD2 and cathelicidins (PG1–5) were concentration-dependently induced by those compounds in both IPEC-J2 and 3D4/31 cells. Furthermore, the antibacterial activities of 3D4/31 cells were augmented following 24 h exposure to HDAC inhibitors. In conclusion, a cell-based high-throughput screening assay was developed for the discovery of porcine HDP inducers, and newly identified HDP-inducing compounds may have potential to be developed as alternatives to antibiotics for applications in swine and possibly other animal species.

Crispene E, a cis-clerodane diterpene inhibits STAT3 dimerization in breast cancer cells

2015 ◽  
Vol 13 (13) ◽  
pp. 3882-3886 ◽  
Author(s):  
Julia Mantaj ◽  
S. M. Abdur Rahman ◽  
Bishwajit Bokshi ◽  
Choudhury M. Hasan ◽  
Paul J. M. Jackson ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Cell Line ◽  
Breast Cancer Cell ◽  
Breast Cancer Cell Line ◽  
Breast Cancer Cells ◽  
Target Genes ◽  
Cancer Cell Line ◽  
Significant Toxicity ◽  
A Cell

Crispene E inhibited STAT3 dimerization in a cell-free fluorescent polarization assay and was found to have significant toxicity against STAT3-dependent MDA-MB 231 breast cancer cell line and selectively inhibited the expression of STAT3 and STAT3 target genes.

Large scale analysis of transcription factor TTF-1/NKX2.1 target genes in GnRH secreting cell line GT1-7

2010 ◽  
Vol 323 (2) ◽  
pp. 215-223 ◽  
Author(s):  
Claudia Provenzano ◽  
Barbara Pascucci ◽  
Eliana Lupari ◽  
Donato Civitareale
Keyword(s):  
Transcription Factor ◽  
Cell Line ◽  
Large Scale ◽  
Target Genes ◽  
Scale Analysis ◽  
Secreting Cell ◽  
Large Scale Analysis

scholarly journals iCSDB: an integrated database of CRISPR screens

2020 ◽  
Vol 49 (D1) ◽  
pp. D956-D961
Author(s):  
Ahyoung Choi ◽  
Insu Jang ◽  
Heewon Han ◽  
Min-Seo Kim ◽  
Jinhyuk Choi ◽  
...  
Keyword(s):  
Cell Lines ◽  
Human Cell ◽  
High Throughput Screening ◽  
Large Scale ◽  
Target Genes ◽  
Human Cell Lines ◽  
Guide Rna ◽  
Functional Studies ◽  
Screening Experiments ◽  
Public Data

Abstract High-throughput screening based on CRISPR-Cas9 libraries has become an attractive and powerful technique to identify target genes for functional studies. However, accessibility of public data is limited due to the lack of user-friendly utilities and up-to-date resources covering experiments from third parties. Here, we describe iCSDB, an integrated database of CRISPR screening experiments using human cell lines. We compiled two major sources of CRISPR-Cas9 screening: the DepMap portal and BioGRID ORCS. DepMap portal itself is an integrated database that includes three large-scale projects of CRISPR screening. We additionally aggregated CRISPR screens from BioGRID ORCS that is a collection of screening results from PubMed articles. Currently, iCSDB contains 1375 genome-wide screens across 976 human cell lines, covering 28 tissues and 70 cancer types. Importantly, the batch effects from different CRISPR libraries were removed and the screening scores were converted into a single metric to estimate the knockout efficiency. Clinical and molecular information were also integrated to help users to select cell lines of interest readily. Furthermore, we have implemented various interactive tools and viewers to facilitate users to choose, examine and compare the screen results both at the gene and guide RNA levels. iCSDB is available at https://www.kobic.re.kr/icsdb/.

Development and characterization of a cell line for large-scale, serum-free production of recombinant adeno-associated viral vectors

2004 ◽  
Vol 6 (12) ◽  
pp. 1369-1381 ◽  
Author(s):  
Deborah Farson ◽  
Thomas C. Harding ◽  
Lucy Tao ◽  
Jun Liu ◽  
Sandra Powell ◽  
...  
Keyword(s):  
Cell Line ◽  
Viral Vectors ◽  
Large Scale ◽  
Serum Free ◽  
A Cell
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