{"doi":"10.1101/2022.08.31.505972","title":"CRISPR Mediated Transactivation in the Human Disease Vector <i>Aedes aegypti</i>","abstract":"Abstract As a major insect vector of multiple arboviruses, Aedes aegypti poses a significant global health and economic burden. A number of genetic engineering tools have been exploited to understand its biology with the goal of reducing its impact. For example, current tools have focused on knocking-down RNA transcripts, inducing loss-of-function mutations or expressing exogenous DNA. However, methods for transactivating endogenous genes have not been developed. To fill this void, here we developed a CRISPR activation (CRISPRa) system in Ae. aegypti to transactivate target gene expression. Gene expression is activated through pairing a catalytically-inactive (‘dead’) Cas9 (dCas9) with a highly-active tripartite activator, VP64-p65-Rta (VPR) and synthetic guide RNA (sgRNA) complementary to a user defined target-gene promoter region. As a proof of concept, we demonstrate that engineered Ae. aegypti mosquitoes harboring a binary CRISPRa system can be used to effectively overexpress two developmental genes, even-skipped (eve) and hedgehog (hh) , resulting in observable morphological phenotypes. We also used this system to overexpress the positive transcriptional regulator of the Toll immune pathway known as AaRel1 , which resulted in a significant suppression of dengue virus serotype 2 (DENV2). This system provides a versatile tool for research pathways not previously possible in Ae. aegypti , such as programmed overexpression of endogenous genes, and may lead to the development of innovative vector control tools.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2022,"id":304197,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9536,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2022-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":566998,"name":"Elena Dalla Benetta","orcid":"0000-0003-2556-8500","position":1,"is_corresponding":false},{"id":325310,"name":"Yuemei Dong","orcid":"0000-0002-7902-2899","position":2,"is_corresponding":false},{"id":996998,"name":"Yunchong Zhao","orcid":"0009-0006-5521-9749","position":3,"is_corresponding":false},{"id":282448,"name":"Ting Yang","orcid":"0000-0001-7201-4231","position":4,"is_corresponding":false},{"id":282256,"name":"Ming Li","orcid":"0000-0002-7578-4968","position":5,"is_corresponding":false},{"id":242214,"name":"Igor Antoshechkin","orcid":"0000-0002-9934-3040","position":6,"is_corresponding":false},{"id":282255,"name":"Anna Buchman","orcid":"0000-0002-8775-6147","position":7,"is_corresponding":false},{"id":887139,"name":"Vanessa Bottino-Rojas","orcid":"0000-0001-8019-9942","position":8,"is_corresponding":false},{"id":245852,"name":"Anthony A. James","orcid":"0000-0001-5577-3308","position":9,"is_corresponding":false},{"id":996999,"name":"Michael Perry","orcid":"0000-0002-5977-8031","position":10,"is_corresponding":false},{"id":111423,"name":"George Dimopoulos","orcid":"0000-0001-6755-8111","position":11,"is_corresponding":false},{"id":262214,"name":"Omar S. Akbari","orcid":"0000-0002-6853-9884","position":12,"is_corresponding":false},{"id":627628,"name":"Michelle Bui","orcid":"0000-0001-9198-8234","position":0,"is_corresponding":true}],"reference_count":62,"raw_metadata":null,"created_at":"2026-07-19T00:32:32.651796Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}