Table 3.
Efficiency of φC31-mediated DNA integration in founder lines
| Founder line | Number of constructs injecteda | Survival rate of microinjected embryos | Integration efficiency |
|---|---|---|---|
| DE-CadGX23w[−]/CyO | 6 | 16.8% (± 3.2%) | 1.4% (± 0.6%) |
| DE-CadGX6w[−]/CyO | 3 | 23.7% (± 4.9%) | 1.3% (± 1.2%) |
| crbGX24w[−]/CyO | 8 | 28.1% (± 3.0%) | 0.7% (± 0.6%) |
| lglGX7w[−]/CyO | 3 | 34.5% (± 3.1%) | 0.9% (± 1.2%) |
| CG31158GX6w[−]/TM3 Sb | 3 | 28.1% (± 1.5%) | 2.9% (± 2.5%) |
| DE-CaGX23w[−]/CyO; vasa-φC31ZH-102D/+ | 30 | 19.4% (± 5.4%) | 7.1% (± 3.4%) |
| crbGX24w[−]/CyO; vasa-φC31ZH-102D/+ | 21 | 28.8% (± 7.5%) | 6.3% (± 2.8%) |
| lglGX7w[−]/CyO; vasa-φC31ZH-102D/+ | 7 | 27.5% (± 8.5%) | 1.1% (± 0.6%) |
| vasa-φC31ZH-2A/vasa-φC3ZH-2A; dArf6GX16w[−]/CyO; | 3 | 7.4% (± 2.5%)b | 9.7% (± 5.4%) |
| sdtGX73w[−]/FM7; vasa-φC31ZH-102D/+ | 1 | 21.7% (± n/a) | 5.6%c (± n/a) |
aThese constructs were based on pGE-attB or pGE-attBGMR (see Table S2).
bThe low survival rate in vasa-φC31ZH-2A/vasa-φC3ZH-2A; dArf6GX16w[−]/CyO could be related to the homozygous copies of vasa-φC31ZH-2A on the X chromosome. We have found that having homozygous copies of vasa-φC31 may adversely affect the healthiness and survival rate of micro-injected embryos of several founder lines, such as DE-CadGX23w[−] and crbGX24w[−], possibly because of certain unique interactions between vasa-φC31 and the attP-50 landing sites or balancers in dArf6, DE-Cad and crb founder lines.
cOnly sdtGX73w[−]/FM7 females were used to set up crosses and to calculate the integration efficiency. FM7/FM7 females and FM7/Y males were discarded since they did not carry the sdtGX73w[−] chromosome.