Table 3.
Successful targeted transgenesis in dual site RosaBxb-GT/GA strains.
| Donor ID | Donor size (kb) | # embryos | RMCE POS | Note | |
|---|---|---|---|---|---|
| B6 background (B6.RosaBxb-GT/GA) | |||||
| P-1 | 1.5 | 78 | 9/23 (39%) | ||
| P-2 | 3.2 | 128 | 1/40 (3%) | ||
| P-3 | 3.3 | 130 | 2/36 (6%) | ||
| P-4 | 3.3 | 130 | 5/36 (14%) | ||
| P-5 | 3.7 | 129 | 16/37 (43%) | ||
| P-6 | 3.7 | 149 | 8/48 (17%) | ||
| P-7 | 4.5 | 63 | 0/11 (0%) | Rescued by upon repeat | |
| P-7 (RPT) | 4.5 | 106 | 3/22 (14%) | ||
| P-8 | 4.6 | 158 | 10/43 (23%) | ||
| P-9 | 5.7 | 165 | 6/50 (12%) | ||
| P-10 | 5.8 | 140 | 7/39 (18%) | ||
| P-11 | 6.8 | 182 | 10/54 (19%) | Krt18-ACE2 | |
| P-12 | 7.1 | 150 | 1/31 (3%) | Improved by upon repeat | |
| P-12 (RPT) | 7.1 | 190 | 12/63 (19%) | ||
| P-13 | 8.5 | 105 | 1/33 (3%) | ||
| P-14 | 8.8 | 62 | 1/14 (7%) | ||
| P-15 | 9.4 | 73 | 0/22 (0%) | Rescued by upon repeat | |
| P-15 (RPT) | 9.4 | 143 | 8/44 (18%) | ||
| P-16 | 10.3 | 67 | 2/11 (18%) | ||
| P-17 | 11.5 | 102 | 1/37 (3%) | ||
| P-18 | 12.3 | 137 | 4/36 (11%) | ||
| P-19 | 13.0 | 115 | 2/21 (10%) | ||
| P-20 | 13.0 | 104 | 1/33 (3%) | ||
| P-21 | 23.6 | 88 | 0/28 (0%) | Rescued by upon repeat | |
| P-21 (RPT) | 23.6 | 107 | 2/26 (8%) | ||
| P-22 | 25.9 | 112 | 2/26 (8%) | ||
| P-23 | 30.6 | 139 | 4/35 (11%) | ||
| P-24 | 42.9 | 129 | 3/21 (14%) | ||
| Total | 1.5–42.9 kb | 3381 | 121/890 (14%) | ||
| NSG background (NSG.RosaBxb-GT/GA) | |||||
| P-11 | 6.8 | 260 | 18/63 (29%) | Krt18-ACE2 | |
| P-25 | 9.1 | 162 | 8/20 (40%) | ||
| Total | 6.8–9.1 kb | 422 | 26/83 (31%) | ||
| All backgrounds combined (B6 and NSG) | |||||
| Total | 1.5–42.9 kb | 3803 | 147/973 (15%) | ||
Summary of 29 independent attempts to integrate 25 unique donor transgenes into the RosaBxb-GT/GA dual-site allele in B6 and/or NSG genetic backgrounds. Shown are DNA donor size, the number of microinjected embryos transferred, and the number of positive Founders animals which carry the transgene, expressed as a percentage of the number of animals screened. The Krt18-ACE2 construct (P-11) was delivered to both B6. RosaBxb-GT/GA and NSG.RosaBxb-GT/GA hosts, highlighted in bold. For a few constructs (P-7, P12, P-15, P-21), the microinjection was repeated following re-purification of the donor vector, which resulted in improved RMCE rate.