Table I. Summary of GT experiments targeting ALS locus
TF FirstTF Second Transformed CalliaBSR CalliW548L, S627I Mutations in Endogenous ALS GeneRatio of Calli with W548L and S627I (%)bRecovery of Regenerated Plantsc
W548L and S627IW548LNone
Exp. 1
pCAMBIA-sGFP Mix2753 (13.7g)30002100
pZH_MMCas9 Mix3994 (15.5g)30002100
Exp. 2
pCAMBIA-sGFP no. 167420002000
pCAMBIA-sGFP no. 1066500000000
pCAMBIA-sGFP no. 1258920002000
pZH_MMCas9 no. 182140002200
pZH_MMCas9 no. 288100000000
pZH_MMCas9 no. 383010001000
Exp. 3
pZH_MMCas9 Mix87220002000
pZH_gLig4-1_MMCas9 Mix98519080740.8124
pZH_gLig4-2_MMCas9 Mix82912131430.4833
Exp. 4
pZH_MMCas9 no. 2110900000000
pZH_MMCas9 no. 39297030310.3233
pZH_gLig4-1_MMCas9 no. 17299030330.4122
pZH_gLig4-1_MMCas9 no. 2110024381841.0007
pZH_gLig4-2_MMCas9 no. 56825010220.1471
pZH_gLig4-2_MMCas9 no. 10100310040510.3992
Exp. 5
T1 (lig4, +1(A) homo; pZH_gLig4-1_MMCas9, segregated out)87700000000
  • a Number of calli used for the transformation of GT vector, pPZ_mALS_g2&3.

  • b GT (%) = (Number of calli with W548L and S627I mutations in endogeneous ALS gene/TF second transformed calli) × 100.

  • c Number of GT lines, in which regenerated plants were obtained.