Insect Biochemistry and Molecular Biology最新文献

{"title":"Functional importance of groups I and II chitinases, CHT5 and CHT10, in turnover of chitinous cuticle during embryo hatching and post-embryonic molting in the red flour beetle, Tribolium castaneum","authors":"Myeongjin Kim ,&nbsp;Mi Young Noh ,&nbsp;Seulgi Mun ,&nbsp;Subbaratnam Muthukrishnan ,&nbsp;Karl J. Kramer ,&nbsp;Yasuyuki Arakane","doi":"10.1016/j.ibmb.2024.104087","DOIUrl":"10.1016/j.ibmb.2024.104087","url":null,"abstract":"<div><p><em>Chitinases</em> (<em>CHT</em>) comprise a large gene family in insects and have been classified into at least eleven subgroups. Many studies involving RNA interference (RNAi) have demonstrated that depletion of group I (CHT5s) and group II (CHT10s) <em>CHT</em> transcripts causes lethal molting arrest in several insect species including the red flour beetle, <em>Tribolium castaneum</em>, presumably due to failure of degradation of chitin in their old cuticle. In this study we investigated the functions of <em>CHT5</em> and <em>CHT10</em> in turnover of chitinous cuticle in <em>T. castaneum</em> during embryonic and post-embryonic molting stages. RNAi and transmission electron microscopic (TEM) analyses indicate that CHT10 is required for cuticular chitin degradation at each molting period analyzed, while CHT5 is essential for pupal-adult molting only. We further analyzed the functions of these genes during embryogenesis in <em>T. castaneum</em>. Real-time qPCR analysis revealed that peak expression of <em>CHT10</em> occurred prior to that of <em>CHT5</em> during embryonic development as has been observed at post-embryonic molting periods in several other insect species. With immunogold-labeling TEM analysis using a fluorescein isothiocyanate-conjugated chitin-binding domain protein (FITC-CBD) probe, chitin was detected in the serosal cuticle but not in any other regions of the eggshell including the chorion and vitelline membrane layers. Injection of double-stranded RNA (dsRNA) for <em>CHT5</em> (ds<em>CHT5</em>), <em>CHT10</em> (ds<em>CHT10</em>) or their co-injection (ds<em>CHT5/10</em>) into mature adult females had no effect on their fecundity and the resulting embryos developed normally inside the egg. There were no obvious differences in the morphology of the outer chorion, inner chorion and vitelline membrane among eggs from these dsRNA-treated females. However, unlike ds<em>CHT5</em> eggs, ds<em>CHT10</em> and ds<em>CHT5/10</em> eggs exhibited failure of turnover of the serosal cuticle in which the horizontal chitinous laminae remained intact, resulting in lethal embryo hatching defects. These results indicate that group I CHT5 is essential for pupal-adult molting, whereas group II CHT10 plays an essential role in cuticular chitin degradation in <em>T. castaneum</em> during both embryonic hatching and all of the post-embryonic molts. CHT10 can serve in place of CHT5 in chitin degradation, except during the pupal-adult molt when both enzymes are indispensable to complete eclosion.</p></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"166 ","pages":"Article 104087"},"PeriodicalIF":3.8,"publicationDate":"2024-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139649039","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Drosophila cytokine GBP2 exerts immune responses and regulates GBP1 expression through GPCR receptor Mthl10","authors":"Masaya Ono ,&nbsp;Takashi Matsumura ,&nbsp;Eui Jae Sung ,&nbsp;Takashi Koyama ,&nbsp;Masanori Ochiai ,&nbsp;Stephen B. Shears ,&nbsp;Yoichi Hayakawa","doi":"10.1016/j.ibmb.2024.104086","DOIUrl":"10.1016/j.ibmb.2024.104086","url":null,"abstract":"<div><p>Growth-blocking peptide (GBP), an insect cytokine, was first found in armyworm <em>Mythimna separata</em>. A functional analogue of GBP, stress-responsive peptide (SRP), was also identified in the same species. SRP gene expression has been demonstrated to be enhanced by GBP, indicating that both cytokines are organized within a hierarchical regulatory network. Although GBP1 (CG15917) and GBP2 (CG11395) have been identified in <em>Drosophila melanogaster</em>, immunological functions have only been characterized for GBP1. It is expected that the biological responses of two structurally similar peptides should be coordinated, but there is little information on this topic. Here, we demonstrate that GBP2 replicates the GBP1-mediated cellular immune response from <em>Drosophila</em> S2 cells. Moreover, the GBP2-induced response was silenced by pre-treatment with dsRNA targeting the GBP receptor gene, <em>Mthl10</em>. Furthermore, treatment of S2 cells with GBP2 enhanced <em>GBP1</em> expression levels, but GBP1 did not affect <em>GBP2</em> expression. GBP2 derived enhancement of <em>GBP1</em> expression was not observed in the presence of GBP1, indicating that GBP2 is an upstream expressional regulator of a GBP1/GBP2 cytokine network. GBP2-induced enhancement of <em>GBP1</em> expression was not observed in <em>Mthl10</em> knockdown cells. Enhancement of <em>GBP2</em> expression was observed in both <em>Drosophila</em> larvae and S2 cells under heat stress conditions; expressional enhancement of both <em>GBP1</em> and <em>GBP2</em> was eliminated in <em>Mthl10</em> knockdown cells and larvae. Finally, Ca²⁺ mobilization assay in GCaMP3-expressing S2 cells demonstrated that GBP2 mobilizes Ca²⁺ upstream of Mthl10. Our finding revealed that <em>Drosophila</em> GBP1 and GBP2 control immune responses as well as their own expression levels through a hierarchical cytokine network, indicating that <em>Drosophila</em> GBP1/GBP2 system can be a simple model that is useful to investigate the detailed regulatory mechanism of related cytokine complexes.</p></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"167 ","pages":"Article 104086"},"PeriodicalIF":3.8,"publicationDate":"2024-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139590144","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bmpali, Bmb1 and Bmcap are necessary for uric acid granule formation in Bombyx mori","authors":"Linmeng Tang ,&nbsp;Dongbin Chen ,&nbsp;Dehong Yang ,&nbsp;Zhiwei Liu ,&nbsp;Xu Yang ,&nbsp;Yujia Liu ,&nbsp;Liying Zhang ,&nbsp;Zulian Liu ,&nbsp;Yaohui Wang ,&nbsp;Zheng Tang ,&nbsp;Yongping Huang","doi":"10.1016/j.ibmb.2024.104075","DOIUrl":"10.1016/j.ibmb.2024.104075","url":null,"abstract":"<div><p>Uric acid is the end-product of nitrogen metabolism of the silkworm and other lepidopterans. The accumulation of uric acid particles in the epidermis causes the larval silkworm to appear white and opaque. However, the mechanism of uric acid granule formation is still unclear. Silkworm epidermis color is linked to the genes responsible for uric acid particle formation. We first identified two genes in the <em>Bombyx mori</em> genome that encode subunits of the Bloc-1 (Biogenesis of Lysosome-related Organelles Complex-1) by homology to these genes in other eukaryotes, <em>Bmpali</em> and <em>Bmb1</em>. Mutation in these genes caused a transparent phenotype in the silkworm larvae, and the loss of BmBloc-1 subunit gene <em>Bmcap</em> resulted in the same phenotype. These three genes are highly conserved between human and silkworm. We discovered that <em>Bmpali</em>, <em>Bmcap</em>, and <em>Bmb1</em> localize in the cytoplasm of BmN cells. Yeast two-hybrid assays demonstrated that the Bmpali physically interacts with both Bmcap and Bmb1. Investigating the roles of <em>Bmpali</em>, <em>Bmb1</em>, and <em>Bmcap</em> is essential for uric acid granule formation understanding in <em>Bombyx mori</em>. These mutants present a valuable silkworm model for studying the biogenesis of lysosome-related organelles (LROs).</p></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"167 ","pages":"Article 104075"},"PeriodicalIF":3.8,"publicationDate":"2024-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139566470","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metabolic and transcriptomic characterization of summer and winter dormancy in the solitary bee, Osmia lignaria","authors":"Dacotah Melicher ,&nbsp;Alex S. Torson ,&nbsp;George D. Yocum ,&nbsp;Jordi Bosch ,&nbsp;William P. Kemp ,&nbsp;Julia H. Bowsher ,&nbsp;Joseph P. Rinehart","doi":"10.1016/j.ibmb.2024.104074","DOIUrl":"10.1016/j.ibmb.2024.104074","url":null,"abstract":"<div><p>The solitary bee <em>Osmia lignaria</em> is a native pollinator in North America with growing economic importance. The life cycle of <em>O. lignaria</em> provides a unique opportunity to compare the physiological and molecular mechanisms underlying two ecologically contrasting dormancies within the same species. <em>O. lignaria</em> prepupae become dormant during the summer to avoid high temperatures. Shortly after adult eclosion, they enter a second dormancy and overwinter as diapausing adults. To compare these two dormancies, we measured metabolic rates and gene expression across development as bees initiate, maintain, and terminate both prepupal (summer) and adult (overwintering) dormancies. We observed a moderate temperature-independent decrease in gas exchange during both the prepupal dormancy after cocoon spinning (45 %) and during adult diapause after eclosion (60 %). We sequenced and assembled a high-quality reference genome from a single haploid male bee with a contiguous n50 of 5.5 Mbp to facilitate our transcriptomic analysis. The transcriptomes of dormant prepupae and diapausing adults clustered into distinct groups more closely associated with life stage than dormancy status. Membrane transport, membrane-bound cellular components, oxidoreductase activity, glutathione metabolism, and transcription factor activity increased during adult diapause, relative to prepupal dormancy. Further, the transcriptomes of adults in diapause clustered into two groups, supporting multiple phases of diapause during winter. Late adult diapause was associated with gene expression profiles supporting increased insulin/IGF, juvenile hormone, and ecdysone signaling.</p></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"166 ","pages":"Article 104074"},"PeriodicalIF":3.8,"publicationDate":"2024-01-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0965174824000055/pdfft?md5=f9a7801d49b0e6ce8208b8eb94f0e4c3&pid=1-s2.0-S0965174824000055-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139469290","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A single mutation in the mosquito (Aedes aegypti) olfactory receptor 8 causes loss of function to 1-octen-3-ol","authors":"Olga Frunze,&nbsp;Dain Lee,&nbsp;Seungha Lee,&nbsp;Hyung Wook Kwon","doi":"10.1016/j.ibmb.2023.104069","DOIUrl":"10.1016/j.ibmb.2023.104069","url":null,"abstract":"<div><p>The host-seeking behavior of mosquitoes have long been established to be primarily odor-mediated. In this process, olfactory receptors (Ors) play a critical role. 1-Octen-3-ol is a common volatile compound that is attractive to hematophagous arthropods such as mosquitos. The olfactory receptor 8 (AaOr8) on the tip of the stylet and maxillary palp of <em>Aedes aegypti</em> is tuned to 1-octen-3-ol, which is required for mosquitoes to quickly find blood vessels from a vertebrate host. However, little is known about the interaction of AaOr8 with 1-octen-3-ol which was studied <em>in vivo</em> and <em>in silico</em> in this study. The molecular binding poses and energies between ligands and the receptor were investigated. Three mutants of AaOr8 were cloned and compared with <em>in vivo</em> calcium imaging utilizing heterologous expression systems. As a result, our findings imply that a genetic disruption including targeted modification of <em>Or</em>s genes may be used to reduce mosquito bites.</p></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"167 ","pages":"Article 104069"},"PeriodicalIF":3.8,"publicationDate":"2024-01-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139465724","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Peritrophins are involved in the defense against Bacillus thuringiensis and nucleopolyhedrovirus formulations in Spodoptera littoralis (Lepidoptera: Noctuidae)","authors":"Gözde Güney ,&nbsp;Doga Cedden ,&nbsp;Sabine Hänniger ,&nbsp;Dwayne D. Hegedus ,&nbsp;David G. Heckel ,&nbsp;Umut Toprak","doi":"10.1016/j.ibmb.2024.104073","DOIUrl":"10.1016/j.ibmb.2024.104073","url":null,"abstract":"<div><p><span><span><span>The peritrophic matrix (or peritrophic membrane, PM) is present in most insects where it acts as a barrier to mechanical insults and pathogens, as well as a facilitator of digestive processes. The PM is formed by the binding of structural </span>PM proteins<span>, referred to as peritrophins, to chitin fibrils and spans the entire midgut in </span></span>lepidopterans. To investigate the role of peritrophins in a highly polyphagous lepidopteran pest, namely the cotton leafworm (</span><span><em>Spodoptera littoralis</em></span>), we generated Insect Intestinal Mucin (IIM⁻) and non-mucin Peritrophin (PER⁻<span>) mutant strains via CRISPR/Cas9 mutagenesis. Both strains exhibited deformed PMs and retarded developmental rates. Bioassays conducted with </span><span><em>Bacillus thuringiensis</em></span> (<em>Bt</em><span>) and nucleopolyhedrovirus (SpliNPV) formulations showed that both the IIM</span>⁻ and PER⁻<span> mutant larvae were more susceptible to these bioinsecticides compared to the wild-type (WT) larvae with intact PM. Interestingly, the provision of chitin-binding agent Calcofluor (CF) in the diet lowered the toxicity of </span><em>Bt</em> formulations in both WT and IIM⁻ larvae and the protective effect of CF was significantly lower in PER⁻ larvae. This suggested that the interaction of CF with PER is responsible for <em>Bt</em> resistance mediated by CF. In contrast, the provision of CF caused increased susceptibility to SpliNPV in both mutants and WT larvae. The study showed the importance of peritrophins in the defense against pathogens in <em>S. littoralis</em> and revealed novel insights into CF-mediated resistance to Cry toxin.</p></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"166 ","pages":"Article 104073"},"PeriodicalIF":3.8,"publicationDate":"2024-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139423978","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"From the fat body to the hemolymph: Profiling tick immune and storage proteins through transcriptomics and proteomics","authors":"Veronika Urbanová ,&nbsp;Stephen Lu ,&nbsp;Eliška Kalinová ,&nbsp;Larissa Martins ,&nbsp;Tereza Kozelková ,&nbsp;Filip Dyčka ,&nbsp;José M. Ribeiro ,&nbsp;Ondřej Hajdušek ,&nbsp;Jan Perner ,&nbsp;Petr Kopáček","doi":"10.1016/j.ibmb.2024.104072","DOIUrl":"10.1016/j.ibmb.2024.104072","url":null,"abstract":"<div><p><span>Ticks are blood-feeding arachnids that are known to transmit various pathogenic microorganisms to their hosts. During blood feeding, ticks activate their metabolism and immune system to efficiently utilise nutrients from the host's blood and complete the feeding process. In contrast to insects, in which the fat body is known to be a central organ that controls essential metabolic processes and immune defense mechanisms, the function of the fat body in tick physiology is still relatively unexplored. To fill this gap, we sought to uncover the repertoire of genes expressed in the fat body associated with trachea (FB/Tr) by analyzing the transcriptome of individual, partially fed (previtellogenic) </span><span><em>Ixodes ricinus</em></span><span><span><span> females. The resulting catalog of individual mRNA sequences<span> reveals a broad repertoire of transcripts encoding proteins involved in nutrient storage and distribution, as well as components of the tick immune system. To gain a detailed insight into the secretory products of FB/Tr specifically involved in inter-tissue transport and humoral immunity, the </span></span>transcriptomic data were complemented with the </span>proteome<span><span> of soluble proteins in the hemolymph of partially fed female ticks. Among these proteins, the hemolipoglyco-carrier proteins were predominant. When comparing immune peptides and proteins from the fat body with those produced by </span>hemocytes, we found that the fat body serves as a unique producer of certain immune components. Finally, time-resolved transcriptional regulation of selected immune transcripts from the FB/Tr was examined in response to experimental challenges with model microbes and analyzed by RT-qPCR. Overall, our data show that the fat body of ticks, similar to insects, is an important metabolic tissue that also plays a remarkable role in immune defense against invading microbes. These findings improve our understanding of tick biology and its impact on the transmission of tick-borne pathogens.</span></span></p></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"165 ","pages":"Article 104072"},"PeriodicalIF":3.8,"publicationDate":"2024-01-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139104718","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The biology of insect chitinases and their roles at chitinous cuticles","authors":"Dhyeykumar Rabadiya,&nbsp;Matthias Behr","doi":"10.1016/j.ibmb.2024.104071","DOIUrl":"10.1016/j.ibmb.2024.104071","url":null,"abstract":"<div><p>Chitin is one of the most prevalent biomaterials in the natural world. The chitin matrix formation and turnover involve several enzymes for chitin synthesis, maturation, and degradation. Sequencing of the <em>Drosophila</em> genome more than twenty years ago revealed that insect genomes contain a number of chitinases, but why insects need so many different chitinases was unclear. Here, we focus on insect GH18 family chitinases and discuss their participation in chitin matrix formation and degradation. We describe their variations in terms of temporal and spatial expression patterns, molecular function, and physiological consequences at chitinous cuticles. We further provide insight into the catalytic mechanisms by discussing chitinase protein domain structures, substrate binding, and enzymatic activities with respect to structural analysis of the enzymatic GH18 domain, substrate-binding cleft, and characteristic TIM-barrel structure.</p></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"165 ","pages":"Article 104071"},"PeriodicalIF":3.8,"publicationDate":"2024-01-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S096517482400002X/pdfft?md5=85d3fcfdac5bb345abb11133b0e1419e&pid=1-s2.0-S096517482400002X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139104675","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Aminoacylase efficiently hydrolyses fatty acid amino acid conjugates of Helicoverpa armigera potentially to increase the pool of glutamine","authors":"Gopal S. Kallure ,&nbsp;Shubhranshu Shekhar Sahoo ,&nbsp;Rutuja S. Kale ,&nbsp;Vitthal T. Barvkar ,&nbsp;Ravindar Kontham ,&nbsp;Ashok P. Giri","doi":"10.1016/j.ibmb.2024.104070","DOIUrl":"10.1016/j.ibmb.2024.104070","url":null,"abstract":"<div><p><span>One of the most prevalent bioactive molecules present in the oral secretion (OS) of lepidopteran<span> insects is fatty acid amino acid conjugates (FACs). Insect dietary components have influence on the synthesis and retaining the pool of FACs in the OS. We noted differential and diet-specific accumulation of FACs in the OS of </span></span><span><em>Helicoverpa armigera</em></span><span> by using Liquid Chromatography-Quadrupole Time of Flight Mass Spectrometry<span>. Interestingly, we identified FACs hydrolyzing enzyme aminoacylase (HaACY) in the OS of </span></span><em>H. armigera</em><span><span> through proteomic analysis. Next, we have cloned, expressed, and purified active recombinant HaACY in the bacterial system. Recombinant HaACY </span>hydrolyzes all the six identified FACs in the OS of </span><em>H. armigera</em><span> larvae fed on host and non-host plants and releases respective fatty acid and glutamine. In these six FACs, fatty acid moieties vary while amino acid glutamine was common. Glutamine obtained upon hydrolysis of FACs by HaACY might serve as an amino acid pool for insect growth and development. To understand the substrate choices of HaACY, we chemically synthesized, purified, and characterized all the six FACs. Interestingly, rHaACY also shows hydrolysis of synthetic FACs into respective fatty acid and glutamine. Our results underline the importance of diet on accumulation of FACs and role of aminoacylase(s) in regulating the level of FACs and glutamine.</span></p></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"165 ","pages":"Article 104070"},"PeriodicalIF":3.8,"publicationDate":"2024-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139097030","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"SYNCAS: Efficient CRISPR/Cas9 gene-editing in difficult to transform arthropods","authors":"Sander De Rouck,&nbsp;Antonio Mocchetti,&nbsp;Wannes Dermauw,&nbsp;Thomas Van Leeuwen","doi":"10.1016/j.ibmb.2023.104068","DOIUrl":"10.1016/j.ibmb.2023.104068","url":null,"abstract":"<div><p>The genome editing technique CRISPR/Cas9 has led to major advancements in many research fields and this state-of-the-art tool has proven its use in genetic studies for various arthropods. However, most transformation protocols rely on microinjection of CRISPR/Cas9 components into embryos, a method which is challenging for many species. Alternatively, injections can be performed on adult females, but transformation efficiencies can be very low as was shown for the two-spotted spider mite, <em>Tetranychus urticae</em>, a minute but important chelicerate pest on many crops. In this study, we explored different CRISPR/Cas9 formulations to optimize a maternal injection protocol for <em>T. urticae.</em> We observed a strong synergy between branched amphipathic peptide capsules and saponins, resulting in a significant increase of CRISPR/Cas9 knock-out efficiency, exceeding 20%. This CRISPR/Cas9 formulation, termed SYNCAS, was used to knock-out different <em>T. urticae</em> genes – <em>phytoene desaturase, CYP384A1</em> and <em>Antennapedia</em> – but also allowed to develop a co-CRISPR strategy and facilitated the generation of <em>T. urticae</em> knock-in mutants. In addition, SYNCAS was successfully applied to knock-out <em>white</em> and <em>white-like</em> genes in the western flower thrips, <em>Frankliniella occidentalis</em>. The SYNCAS method allows routine genome editing in these species and can be a game changer for genetic research in other hard to transform arthropods.</p></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"165 ","pages":"Article 104068"},"PeriodicalIF":3.8,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0965174823001625/pdfft?md5=74c32502c71284a2abb7e408e6232986&pid=1-s2.0-S0965174823001625-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139062539","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}