Summary of transgenic founders and lines generated, transgene copy numbers, and γ-globin mRNA expression levels in adult blood calculated as a percentage of total βmaj expression and per copy
Transgenic founder or line by construct . | Copy no.* . | % total γ/βmaj . | % γ/βmaj per copy . | Integration sites . |
|---|---|---|---|---|
| LCRAγ founders or F1 animals | ||||
| A†‡ | 2 | 0.89 | 0.89 | Euchromatic |
| B | 2 | 0 | — | N. telomeric |
| C‡ | 1 | 0 | 0 | Euchromatic |
| D | 1 | 0.1 | — | Telomeric |
| E | > 2 | 1.3 | — | Centromeric |
| F | 1 | 0.1 | — | N. centromeric |
| G‡ | 1 | < 0.1 | < 0.2 | Euchromatic |
| H | ∼ 15 | 10.3 | — | Euchromatic |
| I‡ | 1 | 0.3 | 0.6 | N. centromeric |
| J | > 2 | 0.6 | — | Euchromatic |
| LCR Aγ HPFH-2 lines | ||||
| A | 7 | 25.5 | 7.2 | Telomeric |
| B§ | 4 | 38.5 | 19.2 | Euchromatic |
| C | 1 | 2.8 | 5.6 | Euchromatic |
| D | 3 | 0.8 | 0.52 | N. centromeric |
| E | 10 | 10.2 | 2 | Centromeric |
| F | 5 | 6.6 | 2.64 | Euchromatic |
| G1∥ | 5-6 | 1.8 | 0.6 | Euchromatic# |
| G2∥ | 2 | 0 | 0 | Euchromatic# |
| G3∥ | 1 | 0 | 0 | Euchromatic# |
| LCR Aγ HPFH-3 lines | ||||
| A | 13 | 28.7 | 4.4 | Euchromatic |
| Founder B | See below | 3.5 | — | Euchromatic |
| B1∥ | 4 | 0.4 | 0.2 | Euchromatic |
| B2∥ | ∼ 25 | 10.6 | 0.84 | Euchromatic |
| C | 2 | 0.1 | 0.1 | Euchromatic |
| LCR Aγ HPFH-6 lines | ||||
| Founder¶ | 3 | 28 | — | Euchromatic |
| A | 1 | 0.9 | 1.8 | Euchromatic |
| B | 4 | 0.9 | 0.45 | Telomeric |
| C | 2 | 0.46 | 0.46 | N. centromeric |
Transgenic founder or line by construct . | Copy no.* . | % total γ/βmaj . | % γ/βmaj per copy . | Integration sites . |
|---|---|---|---|---|
| LCRAγ founders or F1 animals | ||||
| A†‡ | 2 | 0.89 | 0.89 | Euchromatic |
| B | 2 | 0 | — | N. telomeric |
| C‡ | 1 | 0 | 0 | Euchromatic |
| D | 1 | 0.1 | — | Telomeric |
| E | > 2 | 1.3 | — | Centromeric |
| F | 1 | 0.1 | — | N. centromeric |
| G‡ | 1 | < 0.1 | < 0.2 | Euchromatic |
| H | ∼ 15 | 10.3 | — | Euchromatic |
| I‡ | 1 | 0.3 | 0.6 | N. centromeric |
| J | > 2 | 0.6 | — | Euchromatic |
| LCR Aγ HPFH-2 lines | ||||
| A | 7 | 25.5 | 7.2 | Telomeric |
| B§ | 4 | 38.5 | 19.2 | Euchromatic |
| C | 1 | 2.8 | 5.6 | Euchromatic |
| D | 3 | 0.8 | 0.52 | N. centromeric |
| E | 10 | 10.2 | 2 | Centromeric |
| F | 5 | 6.6 | 2.64 | Euchromatic |
| G1∥ | 5-6 | 1.8 | 0.6 | Euchromatic# |
| G2∥ | 2 | 0 | 0 | Euchromatic# |
| G3∥ | 1 | 0 | 0 | Euchromatic# |
| LCR Aγ HPFH-3 lines | ||||
| A | 13 | 28.7 | 4.4 | Euchromatic |
| Founder B | See below | 3.5 | — | Euchromatic |
| B1∥ | 4 | 0.4 | 0.2 | Euchromatic |
| B2∥ | ∼ 25 | 10.6 | 0.84 | Euchromatic |
| C | 2 | 0.1 | 0.1 | Euchromatic |
| LCR Aγ HPFH-6 lines | ||||
| Founder¶ | 3 | 28 | — | Euchromatic |
| A | 1 | 0.9 | 1.8 | Euchromatic |
| B | 4 | 0.9 | 0.45 | Telomeric |
| C | 2 | 0.46 | 0.46 | N. centromeric |
The LCR Aγ transgenics assayed were either founders or F1 animals.
N. telomeric indicates near telomeric integration; N. centromeric, near centromeric integration; and —, not determined.
For founders, an estimate of copy number was made by using probes that specifically detect the end fragments of transgenes and determining the ratio of joining versus end fragments that are detected in multicopy head-to-tail arrays. The lack of detection of joining fragments suggested a single transgene copy. Copy numbers for transgenic lines were determined as described in “Materials and methods.”
Rearranged transgene: one copy intact, second copy broken.
F1 generation transgenic animals were analyzed.
Chromosomal rearrangements were observed in this mouse. Transgene integration has occurred in a translocated chromosome. No single copy line could be generated from the multicopy line.
Transgenic lines generated from the same founder because of segregation of independent integration events.
Founder that did not transmit the transgene.
Only one euchromatic integration site was detectable in the founder animal.