Table 3. RVD variants of the original and adapted dTALEs.
dTALE | dTALE RVD | Parental dTALE | Binding affinity score to the LOB1 promoterA | Number of infection cycles for adaptation | Found after 30 infection cyclesC | Detected in replicateD | ||
---|---|---|---|---|---|---|---|---|
1 | 2 | |||||||
ScoreB | Best possible scoreB | |||||||
dTALEWTLOB1 | NI NG NI NI NI HD HD NG HD NG NG NG NG NN HD HD NG NG NN | NAE | 7.35 | 5.34 | NA | NA | NA | |
dTALELBM1 | NI NH NG NN NI NI NG NG NI NH NG NG NG NN HD HD NG NG NN | NA | -F | 5.27 | NA | YES | YES | YES |
dTALELBM2 | NG NI NI NI NI HD HD NG HD NG NG NG NG NN HD HD NG NG NN | NA | 12.87 | 5.34 | NA | NO | NO | NO |
dTALELBM3 | NG NI NG NI NI HD HD NG HD NG NG NG NG NN HD HD NG NG NN | NA | 14.97 | 5.45 | NA | YES | NO | YES |
dTALELBM4 | HD NG HD NI NG NG NG HD NI NI NG NG NG NN HD HD NG NG NN | NA | 21.66 | 5.45 | NA | YES | YES | YES |
dTALELBM5 | NI NG NI NI NI HD HD NG HD NG NG NI NI NN HD HD NG NG NN | NA | 14.17 | 5.14 | NA | YES | YES | NO |
dTALELBM6 | HD HD NG NG NG HD NI NI NG HD NG NG NG NN HD HD NG NG NN | NA | 25.66 | 6.81 | NA | YES | YES | YES |
dTALELBM7 | NI NG NI NG NI NI NI NI HD HD NG NG NG NN HD HD NG NG NN | NA | 16.63 | 5.26 | NA | YES | NO | YES |
Note
A The binding affinity analysis was conducted in LOB1 variant A from sweet orange (Citrus × sinensis). The promoter region was set as the 1,000 bp sequence upstream of the transcriptional start site.
B According to the target finder tool provided by https://tale-nt.cac.cornell.edu/.
C A clone is defined as “detected” if the dTALEs or adapted TALEs were present in plasmids isolated from bacteria at cycle 30. Three independent clones per strain were isolated from Xcc pthA4:Tn5, introduced to E. coli and sequenced.
D Each experiment was conducted with two replicates marked as “1” and “2”. Data states whether the inducted dTALE was identified in each replicate in the duration of the experiment representing both the time of adaptation and the end of the experiment as cycle 30.
E NA: not applicable.
F Score is beyond cutoff.
Blue color indicates adapted TALEs.