Bioinformatic and cell-based tools for pooled CRISPR knockout screening in mosquitos.
Animals
Anopheles
/ genetics
CRISPR-Cas Systems
/ genetics
Cell Line
Computational Biology
/ methods
Gene Knockout Techniques
Gene Library
Genes, Essential
Humans
Mosquito Control
/ methods
Mosquito Vectors
/ genetics
Pest Control, Biological
/ methods
RNA, Guide, Kinetoplastida
/ genetics
Vector Borne Diseases
/ prevention & control
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
24 11 2021
24 11 2021
Historique:
received:
18
04
2021
accepted:
02
11
2021
entrez:
25
11
2021
pubmed:
26
11
2021
medline:
24
12
2021
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 34819517
doi: 10.1038/s41467-021-27129-3
pii: 10.1038/s41467-021-27129-3
pmc: PMC8613219
doi:
Substances chimiques
RNA, Guide
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
6825Subventions
Organisme : NIGMS NIH HHS
ID : P41 GM132087
Pays : United States
Organisme : Howard Hughes Medical Institute
Pays : United States
Commentaires et corrections
Type : ErratumIn
Informations de copyright
© 2021. The Author(s).
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