Electroporation-mediated nucleic acid delivery during non-embryonic stages for gene-function analysis in Anopheles sinensis

Che, Lin-Rong; He, Zhengbo; Liu, Yan; Yan, Zhen‐Tian; Han, Baozhu; Chen, Xiaojie; He, Xingfei; Zhang, Jiajun; Chen, Bin; Qiao, Liang

doi:10.1016/j.ibmb.2020.103500

Cited by 6 publications

(4 citation statements)

References 41 publications

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“…Gene function analysis refers to the prediction, identification and verification of gene function using bioinformatics and different expression systems (Marzorati et al 2021 ), which has been employed to explore metabolic mechanisms inside cells (Che et al 2020 ; Lin et al 2017 ). Gene Ontology (GO) is a comprehensive database describing gene functions, which can be divided into three parts: biological process (BP), cellular component (CC), and molecular function (MF).…”

Section: Resultsmentioning

confidence: 99%

Toxicity and inhibition mechanism of gallic acid on physiology and fermentation performance of Escherichia coli

liu

Bilal

et al. 2022

Bioresour. Bioprocess.

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Gallic acid is a natural phenolic acid that has a stress inhibition effect on Escherichia coli. This study by integrates fermentation characteristics and transcriptional analyses to elucidate the physiological mechanism of E. coli 3110 response to gallic acid. Compared with the control (without stress), the cell growth was severely retarded, and irregular cell morphology appeared in the case of high levels of gallic acid stress. The glucose consumption of E. coli was reduced successively with the increase of gallic acid content in the fermentation medium. After 20 h of gallic acid stress, cofactor levels (ATP, NAD+ and NADH) of E. coli 3110 were similarly decreased, indicating a more potent inhibitory effect of gallic acid on E. coli. The transcriptional analysis revealed that gallic acid altered the gene expression profiles related to five notable differentially regulated pathways. The genes related to the two-component system were up-regulated, while the genes associated with ABC-transporter, energy metabolism, carbon metabolism, and fatty acid biosynthesis were down-regulated. This is the first report to comprehensively assess the toxicity of gallic acid on E. coli. This study has implications for the efficient production of phenolic compounds by E. coli and provides new ideas for the study of microbial tolerance to environmental stress and the identification of associated tolerance targets. Graphical abstract

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Section: Resultsmentioning

confidence: 99%

Toxicity and inhibition mechanism of gallic acid on physiology and fermentation performance of Escherichia coli

liu

Bilal

et al. 2022

Bioresour. Bioprocess.

View full text Add to dashboard Cite

show abstract

“…Gene function analysis refers to the prediction, identi cation and veri cation of gene function using bioinformatics and different expression systems (Marzorati et al 2021), which has been employed to explore metabolic mechanisms inside cells (Che et al 2020;Lin et al 2017). Gene Ontology (GO) is a comprehensive database describing gene functions, which can be divided into three parts: biological process (BP), cellular component (CC), molecular function (MF).…”

Section: Gene Ontology Analysismentioning

confidence: 99%

Toxicity and inhibition mechanism of gallic acid on physiology and fermentation performance of Escherichia coli

liu

Bilal

et al. 2022

Preprint

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Gallic acid is a natural phenolic acid that has a stress inhibition effect on Escherichia coli. This study elucidates the physiological mechanism of E. coli 3110 response to gallic acid by integrating fermentation characteristics and transcriptional analysis. Compared with the control (without stress), the cell growth was severely inhibited, and irregular cell morphology appeared in the case of high levels of gallic acid stress. With the increase of gallic acid content in the fermentation medium, the glucose consumption of E. coli decreased successively. Moreover, cofactor levels (ATP, NAD+ and NADH) of E. coli 3110 were also lowered under gallic acid after 20 h stress, indicating a stronger inhibitory effect of gallic acid on E. coli. The transcriptional analysis revealed that gallic acid altered the gene expression profiles related to five notable differentially regulated pathways. The genes related to the two-component system were up-regulated, while the genes associated with ABC-transporter, energy metabolism, carbon metabolism, and fatty acid biosynthesis were down-regulated. This is the first report on the systematic evaluation of the toxicity of gallic acid on E. coli. This study has a guiding significance for the efficient production of phenolic compounds by E. coli and provides new ideas for the study of microbial tolerance to environmental stress and the discovery of related tolerance targets.

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“…It was first successfully employed in Odonata larvae and later used to facilitate the knockdown of AsCPR128 in An. sinensis [ 15 ]. Both methods may be challenging to scale up for high-throughput testing of gene function or screening for potential vector control targets, although microinjections have been used for this purpose [ 16 ].…”

Section: Introductionmentioning

confidence: 99%

RNAi-Mediated Silencing of Laccase 2 in Culex pipiens Pupae via Dehydration and Soaking Results in Multiple Defects in Cuticular Development

Naumenko,

Fritz

2024

Insects

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Mosquitoes transmit a range of pathogens, causing devastating effects on human health. Population genetic control strategies have been developed and successfully used for several mosquito species. The most important step in identifying potential targets for mosquito control is the understanding of gene function. RNA interference (RNAi) is a powerful tool for gene silencing which has been widely used to study gene function in insects via knockdown of expression. The success of RNAi in insects depends on the efficient delivery of dsRNA into the cells, with microinjections being the most commonly used to study mosquito gene function. However, microinjections in the pupal stage lead to significant mortality in Aedes and Culex species, and few studies have performed microinjections in Culicinae pupae. Advanced techniques, such as CRISPR/Cas9 knockout, require establishing individual mosquito lines for each gene studied, and maintaining such lines may be limited by the insect-rearing capacity of a laboratory. Moreover, at times gene knockout during early development (embryo stage) has a deleterious effect on mosquito development, precluding the analysis of gene function in the pupal and adult stages and its potential for mosquito control. There is a need for a simple procedure that can be used for the fast and reliable examination of adult gene function via RNAi knockdown. Here, we focus on the aquatic stages of the mosquito life cycle and suggest a quick and easy assay for screening the functional role of genes in Culex pipiens mosquitoes without using microinjections. By dehydration of early stage pupae and subsequent rehydration in highly concentrated dsRNA, we achieved a moderate knockdown of laccase 2, a gene that turns on in the pupal stage and is responsible for melanization and sclerotization of the adult cuticle.

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Electroporation-mediated nucleic acid delivery during non-embryonic stages for gene-function analysis in Anopheles sinensis

Cited by 6 publications

References 41 publications

Toxicity and inhibition mechanism of gallic acid on physiology and fermentation performance of Escherichia coli

Toxicity and inhibition mechanism of gallic acid on physiology and fermentation performance of Escherichia coli

Toxicity and inhibition mechanism of gallic acid on physiology and fermentation performance of Escherichia coli

RNAi-Mediated Silencing of Laccase 2 in Culex pipiens Pupae via Dehydration and Soaking Results in Multiple Defects in Cuticular Development

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