{"id":277,"date":"2018-03-07T16:06:51","date_gmt":"2018-03-07T15:06:51","guid":{"rendered":"http:\/\/projects.nib.si\/rnainval\/?page_id=277"},"modified":"2022-08-17T14:22:05","modified_gmt":"2022-08-17T13:22:05","slug":"project-phases-and-realisation","status":"publish","type":"page","link":"https:\/\/projects.nib.si\/rnainval\/project-phases-and-realisation\/","title":{"rendered":"Project phases and realisation"},"content":{"rendered":"

Task 1: Candidate gene selection<\/span><\/p>\n

Realization:\u00a0We have completed all the activities in this task and used the results in Task 2. The genome of the Colorado potato beetle, essential for gene selection, was published in a scientific journal (https:\/\/doi.org\/10.1038\/s41598-018-20154-1<\/a>).<\/p>\n

Task 2:\u00a0Selection of target genes based on expression<\/span><\/p>\n

Realization:\u00a0We have completed all the activities in this task and used the results in Task 3.<\/p>\n

Task 3:\u00a0dsRNA production and delivery method establishment<\/span><\/p>\n

Realization:\u00a0We have completed all the activities in this task and used the delivery method of dsRNA in Task 4.<\/p>\n

Task 4:\u00a0Evaluation of dsRNAs’ insecticidal potential<\/span><\/p>\n

Realization:\u00a0We have completed activities related to the planned laboratory testing, collection, and data processing. The manuscript preprint was uploaded\u00a0to bioRxiv and\u00a0published in\u00a0a scientific journal (https:\/\/doi.org\/10.3389\/fpls.2020.01250<\/a>).<\/p>\n

Hypothesis 1 (suppression of selected candidate genes will result in increased mortality, abnormal development or decreased growth of CPB larvae) was confirmed only for the selected target gene mesh and rejected for the JHBP gene.\u00a0For the\u00a0third selected target gene, the hypothesis could not be confirmed or rejected, since we did not succeed in effectively silencing the expression of the alas gene using the designed dsRNA.<\/p>\n

Task 5:\u00a0dsRNA-induced effects on gut tissue transcriptome using RNA-Seq<\/span><\/p>\n

Realization:\u00a0We have completed activities related to the planned laboratory testing, preparation of bioinformatic works, generation, and data processing. The manuscript\u00a0is in preparation.<\/p>\n

Hypothesis 2 (suppression of genes whose biological function has not yet been confirmed will lead to changes at the level of transcriptome and phenotype associated with the function of the gene) has\u00a0been changed. We have decided to study the effect of non-specific dsRNA on the CPB gut transcriptome instead of the influence of insecticidal dsRNA. The results of this analysis show that the gut responds with a change in expression of about 100 genes. dsRNA thus does not cause major reprogramming of the gut transcriptome. Comparison with the response of Tribolium beetle and\u00a0honey bee to non-specific dsRNA response does\u00a0not suggest that insects would have a general and preserved response to non-specific dsRNA at the level of gene expression.<\/p>\n

Task 6:\u00a0dsRNA-induced effects on gut metagenome<\/span><\/p>\n

Realization:\u00a0We have completed activities related to the planned laboratory testing, preparation of bioinformatic works, generation, and data processing. The time delay from the original plan was due to difficulties in establishing effective isolation of metagenomic DNA from the content of the beetle’s gastrointestinal tract.<\/p>\n

Hypothesis 3 (RNAi insecticide will change the composition of intestinal bacterial clades and accelerate or reduce the abundance of particular clades) has been confirmed by metagenomic analysis. These results will have to be confirmed by qPCR and expanded to samples from the remaining experiments.<\/p>\n","protected":false},"excerpt":{"rendered":"

Task 1: Candidate gene selection Realization:\u00a0We have completed all the activities in this task and used the results in Task 2. The genome of the Colorado potato beetle, essential for gene selection, was published in a scientific journal (https:\/\/doi.org\/10.1038\/s41598-018-20154-1). Task 2:\u00a0Selection of target genes based on expression Realization:\u00a0We have completed all the activities in this […]<\/p>\n","protected":false},"author":4,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"jetpack_post_was_ever_published":false,"footnotes":""},"class_list":["post-277","page","type-page","status-publish","hentry"],"jetpack_sharing_enabled":true,"jetpack_shortlink":"https:\/\/wp.me\/P7OqMV-4t","jetpack-related-posts":[],"jetpack_likes_enabled":true,"_links":{"self":[{"href":"https:\/\/projects.nib.si\/rnainval\/wp-json\/wp\/v2\/pages\/277","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/projects.nib.si\/rnainval\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/projects.nib.si\/rnainval\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/projects.nib.si\/rnainval\/wp-json\/wp\/v2\/users\/4"}],"replies":[{"embeddable":true,"href":"https:\/\/projects.nib.si\/rnainval\/wp-json\/wp\/v2\/comments?post=277"}],"version-history":[{"count":8,"href":"https:\/\/projects.nib.si\/rnainval\/wp-json\/wp\/v2\/pages\/277\/revisions"}],"predecessor-version":[{"id":331,"href":"https:\/\/projects.nib.si\/rnainval\/wp-json\/wp\/v2\/pages\/277\/revisions\/331"}],"wp:attachment":[{"href":"https:\/\/projects.nib.si\/rnainval\/wp-json\/wp\/v2\/media?parent=277"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}