Targeted delivery of CRISPR-Cas9 ribonucleoprotein into arthropod ovaries for heritable germline gene editing

Duverney Chaverra-Rodriguez, Vanessa M. Macias, Grant L. Hughes, Sujit Pujhari, Yasutsugu Suzuki, David R. Peterson, Donghun Kim, Sage McKeand, Jason L. Rasgon

Research output: Contribution to journalArticle

13 Citations (Scopus)

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.

Original languageEnglish (US)
Article number3008
JournalNature Communications
Volume9
Issue number1
DOIs
StatePublished - Dec 1 2018

Fingerprint

Clustered Regularly Interspaced Short Palindromic Repeats
arthropods
editing
ovaries
Ribonucleoproteins
Arthropods
Culicidae
genes
Ovary
delivery
Genes
gametocytes
cargo
peptides
Hemolymph
Microinjections
injection
Oocytes
Peptides
Technology

ASJC Scopus subject areas

  • Chemistry(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Physics and Astronomy(all)

Cite this

Chaverra-Rodriguez, D., Macias, V. M., Hughes, G. L., Pujhari, S., Suzuki, Y., Peterson, D. R., ... Rasgon, J. L. (2018). Targeted delivery of CRISPR-Cas9 ribonucleoprotein into arthropod ovaries for heritable germline gene editing. Nature Communications, 9(1), [3008]. https://doi.org/10.1038/s41467-018-05425-9

Targeted delivery of CRISPR-Cas9 ribonucleoprotein into arthropod ovaries for heritable germline gene editing. / Chaverra-Rodriguez, Duverney; Macias, Vanessa M.; Hughes, Grant L.; Pujhari, Sujit; Suzuki, Yasutsugu; Peterson, David R.; Kim, Donghun; McKeand, Sage; Rasgon, Jason L.

In: Nature Communications, Vol. 9, No. 1, 3008, 01.12.2018.

Research output: Contribution to journalArticle

Chaverra-Rodriguez, D, Macias, VM, Hughes, GL, Pujhari, S, Suzuki, Y, Peterson, DR, Kim, D, McKeand, S & Rasgon, JL 2018, 'Targeted delivery of CRISPR-Cas9 ribonucleoprotein into arthropod ovaries for heritable germline gene editing', Nature Communications, vol. 9, no. 1, 3008. https://doi.org/10.1038/s41467-018-05425-9
Chaverra-Rodriguez, Duverney ; Macias, Vanessa M. ; Hughes, Grant L. ; Pujhari, Sujit ; Suzuki, Yasutsugu ; Peterson, David R. ; Kim, Donghun ; McKeand, Sage ; Rasgon, Jason L. / Targeted delivery of CRISPR-Cas9 ribonucleoprotein into arthropod ovaries for heritable germline gene editing. In: Nature Communications. 2018 ; Vol. 9, No. 1.
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