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Enhanced Golic+: Gene targeting with 100% recovery in Drosophila male germ cells

Hui-Min Chen, Xiaohao Yao, Qingzhong Ren, Chuan-Chie Chang, Ling-Yu Liu, View ORCID ProfileTzumin Lee
doi: https://doi.org/10.1101/681874
Hui-Min Chen
Howard Hughes Medical Institute, Janelia Research Campus, 19700 Helix Drive, Ashburn, VA 20147, USA
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Xiaohao Yao
Howard Hughes Medical Institute, Janelia Research Campus, 19700 Helix Drive, Ashburn, VA 20147, USA
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Qingzhong Ren
Howard Hughes Medical Institute, Janelia Research Campus, 19700 Helix Drive, Ashburn, VA 20147, USA
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Chuan-Chie Chang
Howard Hughes Medical Institute, Janelia Research Campus, 19700 Helix Drive, Ashburn, VA 20147, USA
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Ling-Yu Liu
Howard Hughes Medical Institute, Janelia Research Campus, 19700 Helix Drive, Ashburn, VA 20147, USA
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Tzumin Lee
Howard Hughes Medical Institute, Janelia Research Campus, 19700 Helix Drive, Ashburn, VA 20147, USA
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  • ORCID record for Tzumin Lee
  • For correspondence: leet@janelia.hhmi.org
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ABSTRACT

The efficiency of gene targeting can vary drastically. Even utilizing CRISPR/Cas9 does not ensure rapid, successful gene targeting. In Drosophila, we previously established Golic+ to augment gene-targeting productivity simply through fly pushing. This transgene-based system elicits gene targeting in germ cells. It further implements lethality selection to enrich for progeny with accurate gene targeting. However, limitations still remain. Here we deliver an improved Golic+ technique that we name Enhanced Golic+ (E-Golic+). E-Golic+ incorporates genetic modifications to eliminate false positives while simultaneously boosting efficiency. Strikingly, male germ cells are exceptionally susceptible to gene targeting using E-Golic+. With male founders, we easily achieve 100% recovery of correct gene targeting. Given the superior efficiency/specificity and relatively effortless scalability, E-Golic+ promises to triumph in any challenging and otherwise unattainable gene targeting projects in Drosophila.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted June 24, 2019.
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Enhanced Golic+: Gene targeting with 100% recovery in Drosophila male germ cells
Hui-Min Chen, Xiaohao Yao, Qingzhong Ren, Chuan-Chie Chang, Ling-Yu Liu, Tzumin Lee
bioRxiv 681874; doi: https://doi.org/10.1101/681874
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Enhanced Golic+: Gene targeting with 100% recovery in Drosophila male germ cells
Hui-Min Chen, Xiaohao Yao, Qingzhong Ren, Chuan-Chie Chang, Ling-Yu Liu, Tzumin Lee
bioRxiv 681874; doi: https://doi.org/10.1101/681874

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