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Targeted delivery of CRISPR-Cas9 ribonucleoprotein into arthropod ovaries for heritable germline gene editing.
- Source :
-
Nature communications [Nat Commun] 2018 Aug 01; Vol. 9 (1), pp. 3008. Date of Electronic Publication: 2018 Aug 01. - Publication Year :
- 2018
-
Abstract
- Cas9-mediated gene editing is a powerful tool for addressing research questions in arthropods. Current approaches rely upon delivering Cas9 ribonucleoprotein (RNP) complex by embryonic microinjection, which is challenging, is limited to a small number of species, and is inefficient even in optimized taxa. Here we develop a technology termed Receptor-Mediated Ovary Transduction of Cargo (ReMOT Control) to deliver Cas9 RNP to the arthropod germline by injection into adult female mosquitoes. We identify a peptide (P2C) that mediates transduction of Cas9 RNP from the female hemolymph to the developing mosquito oocytes, resulting in heritable gene editing of the offspring with efficiency as high as 0.3 mutants per injected mosquito. We demonstrate that P2C functions in six mosquito species. Identification of taxa-specific ovary-specific ligand-receptor pairs may further extend the use of ReMOT Control for gene editing in novel species.
- Subjects :
- Alleles
Animals
Base Sequence
Crosses, Genetic
Culicidae embryology
Drosophila melanogaster genetics
Embryo, Nonmammalian metabolism
Female
Green Fluorescent Proteins metabolism
Inheritance Patterns genetics
Injections
Male
Mutation genetics
Oocytes metabolism
Sequence Deletion
CRISPR-Associated Protein 9 metabolism
CRISPR-Cas Systems genetics
Culicidae genetics
Gene Editing
Germ Cells metabolism
Ovary metabolism
Ribonucleoproteins metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 2041-1723
- Volume :
- 9
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Nature communications
- Publication Type :
- Academic Journal
- Accession number :
- 30068905
- Full Text :
- https://doi.org/10.1038/s41467-018-05425-9