Table 1. Summary of targeted integration.
| Target locus | Percentage of mCherry+/ZsGreen1-DR- populationa (%) | Total clone number (mCherry+/ZsGreen1-DR-) | 5′/3′ Junction PCR positive | Out/Out PCR positivec | Targeting efficiencyd (%) |
|---|---|---|---|---|---|
| COSMC | 45.7 ± 2.6 | 115 | 83 | 70 | 27.8 |
| Mgat1b (Before RCA-I enrichment) | 62.2 ± 2.7/41.1 ± 1.0 | 124/37 | 0/0 | N/A | N/A |
| Mgat1b (After RCA-I enrichment) | 35.3 ± 2.1/60.4 ± 6.6 | 31/59 | 10/16 | 9/16 | 10.2/16.4 |
| LdhA | 45.2 ± 2.9 | 61 | 10 | N/A | 7.4 |
aThe numbers indicate the percentage of mCherry+/ZsGreen1-DR- cell population in each stable cell pool measured by imaging cell cytometer. Average values from three independent experiments ± SD are shown.
bTwo numbers separated by diagonal lines indicate the results from two sgRNA target sites (sgRNA1/sgRNA5).
cPositive indicates expected size of PCR products occurring by targeted integration.
dTargeting efficiency was obtained by multiplying the percentage of mCherry+/ZsGreen1-DR- cell population by the percentage of out/out PCR positive clones amongst total mCherry+/ZsGreen1-DR-clones investigated. In case of LdhA locus, out/out PCR results were not available. Therefore, 5′/3′ junction PCR results were used instead.