A platform for the transgenic modification of Sandflies (Lutzomyia longipalpis) and Triatomine bugs (Rhodnius prolixus) towards deriving insect vectors refractory to disease agents mediated by CRISPR-Cas9 gene drive (360G-Wellcome-208913_Z_17_Z)

Vector borne pathogens Leishmania, and Trypanosoma cruzi cause devastating human disease (Leishmaniasis and Chagas disease respectively). We contribute to a new strategy by genetically modifying insect vectors, sandflies (Lutzomyia longipalpis) and triatomine bugs (Rhodnius prolixus), leading eventually to transgenic insects resistant to disease agents. We will use two different transgenic approaches, piggyBac and CRISPR-Cas9. PiggyBac is a method for semi random insertion of exogenous DNA into the genome. CRISPR-Cas9 is a new and highly targeted method to manipulate genes in a manner not previously possible. Microinjection of insect embryos with piggyBac plasmids to obtain insects expressing fluorescent markers for subsequent CRISPR-Cas9 knockout experiments facilitating easy monitoring of outcomes. Knockout of fluorescent markers (above), using CRISPR-Cas9 constructs, demonstrating that system components are functional and that we can precisely target genes of interest. Knockout of endogenous genes by CRISPR-Cas 9 that are non-essential or important in sustaining infection in insect vectors. Insertion and expression of exogenous DNA (fluoresence markers) using CRISPR-Cas9 harnessing homology directed repair facilitating inheritance to offspring at levels above mendelian inheritance (gene drive). Pilot outputs offer real prospects for developing powerful approaches of interrupting disease transmission.

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Grant Details

Amount Awarded 75106
Applicant Surname Yeo
Approval Committee Science Seeds Advisory Panel
Award Date 2017-09-05T00:00:00+00:00
Financial Year 2016/17
Grant Programme: Title Seed Award in Science
Internal ID 208913/Z/17/Z
Lead Applicant Dr Matthew Yeo
Partnership Value 75106
Planned Dates: End Date 2022-03-14T00:00:00+00:00
Planned Dates: Start Date 2018-03-15T00:00:00+00:00
Recipient Org: Country United Kingdom
Region Greater London