Intronic gRNAs for the Construction of Minimal Gene Drive Systems

Frontiers in Bioengineering and Biotechnology

Phillipos Aris Papathanos¹  Astrid Hoermann²  Alexander Nash²  Nikolai Windbichler²  Paolo Capriotti² 

[1] Department of Entomology, Robert H. Smith Faculty of Agriculture, Food and Environment, Hebrew University of Jerusalem, Rehovot, Israel;Department of Life Sciences, Imperial College London, London, United Kingdom;

关键词: gene drives;    genetic control;    Drosophila;    synthetic biology;    genetics;   

DOI  :  10.3389/fbioe.2022.857460

来源: DOAJ

【 摘 要 】

Gene drives are promising tools for the genetic control of insect vector or pest populations. CRISPR-based gene drives are generally highly complex synthetic constructs consisting of multiple transgenes and their respective regulatory elements. This complicates the generation of new gene drives and the testing of the behavior of their constituent functional modules. Here, we explored the minimal genetic components needed to constitute autonomous gene drives in Drosophila melanogaster. We first designed intronic gRNAs that can be located directly within coding transgene sequences and tested their functions in cell lines. We then integrated a Cas9 open reading frame hosting such an intronic gRNA within the Drosophila rcd-1r locus that drives the expression in the male and female germlines. We showed that upon removal of the fluorescent transformation marker, the rcd-1rd allele supports efficient gene drive. We assessed the propensity of this driver, designed to be neutral with regards to fitness and host gene function, to propagate in caged fly populations. Because of their simplicity, such integral gene drives could enable the modularization of drive and effector functions. We also discussed the possible biosafety implications of minimal and possibly recoded gene drives.

【 授权许可】

Unknown