Molecular safeguarding of CRISPR gene drive experiments
- Cornell University, United States
Figures
Figure 1
Schematic diagram of our synthetic target site drive and split drive constructs.
(a) The synthetic target site drive constructs contain Cas9 with the germline nanos promoter and 3’UTR, a dsRed marker with a slightly recoded (*) 3xP3 promoter and P10 3’UTR, and a gRNA driven by the U6:3 promoter that targets the synthetic EGFP gene. The two homology arms include the EGFP sequence with its 3xP3 promoter and SV40 3’UTR regions. (b) The split drive contains a dsRed marker gene driven by a 3xP3 promoter together with a SV40 3’UTR, a gRNA expressed by the U6:3 promoter that targets yellow, and two homology arms for yellow. The unlinked supporting element contains Cas9 driven by the nanos promoter with a nanos 3’UTR, and an EGFP marker gene driven by a 3xP3 promoter together with a SV40 3’UTR.
Tables
Table 1
Drive performances of synthetic target site and split drives compared with the standard drives from our previous studies (Champer et al., 2017; Champer et al., 2018b).
https://doi.org/10.7554/eLife.41439.004| Drive | Male drive conversion efficiency | Female drive conversion efficiency | Embryo r2 resistance rate |
|---|---|---|---|
| EGFP site B | 32 ± 3% | 52 ± 3% | 88 ± 1% |
| EGFP site E | 46 ± 4% | 54 ± 5% | 91 ± 2% |
| EGFP site Y | N/A | 53 ± 3% | 80 ± 2% |
| cinnabar | 39 ± 3% | 54 ± 4% | 100 ± 0% |
| white | N/A | 59 ± 2% | 77 ± 2% |
| Split-yellow | N/A | 74 ± 2% | 74 ± 2% |
| yellow | N/A | 63 ± 3% | 20 ± 2% |
Additional files
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Supplementary file 1
Plasmid construction details and oligonucleotide sequences.
- https://doi.org/10.7554/eLife.41439.005
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Supplementary file 2
Fly phenotype data and rate calculations.
- https://doi.org/10.7554/eLife.41439.006
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Transparent reporting form
- https://doi.org/10.7554/eLife.41439.007
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Molecular safeguarding of CRISPR gene drive experiments
eLife 8:e41439.
https://doi.org/10.7554/eLife.41439
