Table 1.
Size of indels in respective line analysed with Sanger sequencing and HRFA (in brackets), where “0” indicates the wild type allele fragment size, “–” represents a deletion and “ + ” represents an insertion.
| Group | Line | Size of indels confirmed by Sanger sequencing (results from HRFA in brackets) | |
|---|---|---|---|
| Sbe1 | Sbe2 | ||
| Group 1 | 82007 | -38a/-3¥,a/ + 70a (-38/-3/ + 70) | 0¤ (0) |
| 82050 | -22a/-1a/ + 38a/ + 165¥,a,e (-22/ -1/ + 38/ + 166) | 0¤ (0) | |
| 82079 | -1a/ + 47a,e (-1/ + 47) | 0¤ (0) | |
| 104011 | -94a,b/-4a (-95/-4) | 0¤ (0) | |
| 104032 | -55a,b/ + 26a,b (-56/ + 26) | 0¤ (0) | |
| Group 2 | 104001 | -94a,b / + 3¥,a,b (-95/ + 3) | -129¥,c,d/-3¥,c,d/-1c/0¤ (-130/-3/-1/0) |
| 104005 | -93¥,a,b/-92,a,b (-94/-93) | -10d/-4c,d/0¤ (-10/-4/0) | |
| 104006 | -92a,b/-5a,b/-1a (-92/-5/-1) | -11d/-1 d/0¤ (-11/-1/0) | |
| 104016 | -93¥,a,b/-4a,b (-94/-4) | -2c/0¤/ + 6¥,d (-2/0/ + 6) | |
| 104018 | -93¥,a,b/-23a,b/-17a/ + 153¥,a,b (-94/-23/-17/ + 153) | -1c/0¤/ + 104c,d (-1/0/ + 105) | |
| 104034 | -94a,b /-93¥,a,b /-5a,b / + 60¥,a,b (-95/-94/-5/ + 60) | -8c,d/-6¥,c,d/0¤ (-8/-6/0) | |
| Group 3 | 104010 | -5a,b/ + 92a,b/ + 123¥,a,b (-5/ + 93/ + 122) | -2d/-1c/ + 48¥,c,d/ + 194c,d (-2/-1/ + 48/ + 195) |
| 104023 | -92a,b/-3¥,a,b/ + 13a,b / + 251a,b (-92/-3/ + 13/ + 252) | -127c,d/-9¥,c,d/-5c,d (-128/-9/-5) | |
Less than four different indels indicates that at least two alleles share the same genetic context. Lack of “0” means no wild type allele remaining in the line. Group 1 represents lines mutated in all four alleles of Sbe1; Group 2 represents lines mutated in all four alleles of Sbe1 and two to three alleles in Sbe2; Group 3 represents lines mutated in all alleles of both Sbe1 and Sbe2. ¤ Wild type alleles. ¥ In-frame indels. a Indels at BE1T3 target site, b Indels at BE1T4 target site, c Indels at BE2T3 target site and d Indels at BE2T4 target site. e Insert origination from vector used for in vitro transcription.