Table 2. Summary of zebrafish GeneWeld GTag integrations.
| Genomic target | # of germline transmitting adults | # of precise 5’ junctions by PCR | # of precise 3’ junctions by PCR | # of precise integrations by Southern |
|---|---|---|---|---|
| noto | 3/5 | 8/8 | 0/3 and n/d | 0/2 |
| tyr | 3/8 | 1/1 | 1/1 | 1/1 |
| esama | 12/18 | 8/10* | 9/10* | n/d |
| flna | 3/4 | 4/4 | n/d | n/d |
| msna | 1/4 | 12/12 | 1/12 | n/d |
| aqp1a1 | 2/9 | 1/1 | 1/1 | n/d |
| aqp8a1 | 1/1 | 1/1 | 1/1 | n/d |
| Total | 25/49 (51%) | 35/37 (95%) | 13/28 (46%) | 1/3 (33%) |
F1 or F2 embryos were analyzed for junction fragments.
*Embryos from a single esama F0 founder inherited a mix of precise and imprecise junction alleles. Multiple positive FI embryos were obtained in which at least one of the embryos contained precise junctions. A polymorphism in the homology domain was also detected in the esama 5’ junction from F0 #4. One of the F1s from F0#5 also contained an imprecise junction at the 5’ end. esama F1 3’ junctions all contain a single nucleotide variant in the homology arm. Interestingly, this was corrected to the genomic sequence. One esama F1 3’ junction also included a 20 bp insertion.
n/d – not determined.