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• W3217293567 abstract "Mosquito-borne diseases present a worldwide public health burden. Current efforts to understand and counteract them have been aided by the use of cultured mosquito cells. Moreover, application in mammalian cells of forward genetic approaches such as CRISPR screens have identified essential genes and genes required for host-pathogen interactions, and in general, aided in functional annotation of genes. An equivalent approach for genetic screening of mosquito cell lines has been lacking. To develop such an approach, we design a new bioinformatic portal for sgRNA library design in several mosquito genomes, engineer mosquito cell lines to express Cas9 and accept sgRNA at scale, and identify optimal promoters for sgRNA expression in several mosquito species. We then optimize a recombination-mediated cassette exchange system to deliver CRISPR sgRNA and perform pooled CRISPR screens in an Anopheles cell line. Altogether, we provide a platform for high-throughput genome-scale screening in cell lines from disease vector species." @default.
• W3217293567 created "2021-12-06" @default.
• W3217293567 creator A5000699019 @default.
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• W3217293567 date "2021-11-24" @default.
• W3217293567 modified "2023-09-27" @default.
• W3217293567 title "Bioinformatic and cell-based tools for pooled CRISPR knockout screening in mosquitos" @default.
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• W3217293567 doi "https://doi.org/10.1038/s41467-021-27129-3" @default.
• W3217293567 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/8613219" @default.
• W3217293567 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/34819517" @default.
• W3217293567 hasPublicationYear "2021" @default.
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