Table 1.
CRISPR/Cas9 induced LhCd and LhCn mutations in Lygus hesperus.
| Gene | Namea | Mutation corresponding to sgRNA target sites | Typee | gDNA | cDNA | |||
|---|---|---|---|---|---|---|---|---|
| sgRNA 1bc | sgRNA 2bd | Gn | # of samples (# of clones) | Gn | # of samples (# of clones) | |||
| LhCd | ||||||||
| Cd1.1 | 664_667del | 677_696del | ps | 0, 3 | 1 (1), 2 (11) | ndf | nd | |
| Cd1.2 | 666_697delinsCGGGAATTGGATTGAATGC | *g | InF | nd | nd | 2, 3 | 1 (1), 1 (4) | |
| Cd1.3 | 666_668del | 693A > T | InF | 1 | 1 (5) | 1, 2 | 1 (1), 2 (4) | |
| Cd1.4 | 666_670del | 690_723delinsCCGAGTA | ps | 3 | 1 (6) | 2 | 5 (9) | |
| Cd1.5 | 666_669del | 683_697delinsGCACGCAACATGGCTGGCAA | ps | 1 | 1 (5) | 1 | 2 (5) | |
| Cd1.6 | 664_666del | 690_692delinsCTCTGT | InF | 0 | 1 (1) | ndg | nd | |
| Cd1.7 | wth | 692del | ps | 0 | 1 (1) | nd | nd | |
| Cd1.8 | 649_665delinsj | 680_692delinsG | nd | 0 | 1 (1) | nd | nd | |
| Cd1.9 | 666_692delinsGG | * | ps | 0 | 1 (1) | nd | nd | |
| Cd1.10 | 666_670del | 688_693del | ps | 0 | 1 (1) | nd | nd | |
| Cd1.11 | 666_670del | 689_695del | ps | 0 | 1 (1) | nd | nd | |
| Cd1.12 | 649_665delinsTGGGTTTTCCCGAAGGC | 670_693delinsCTCACGAGCTCCCTCCTACCCCTT | InF | 0 | 1 (1) | nd | nd | |
| Cd1.13 | 666_670del | 690_695del | ps | 0 | 1 (1) | nd | nd | |
| Cd1.14 | 666_694delinsGACGCAAGGACG | * | ps | 2 | 1 (6) | nd | nd | |
| Cd1.15 | 664_669delinsTTTG | 693_694insA | ps | nd | nd | 1 | 1 (2) | |
| Cd1.16 | 659_668delinsTCGTTAGTGTTG | 684_697delinsAC | InF | nd | nd | 1 | 1 (2) | |
| Cd1.17 | 666_693delinsTTGCGGGAATTGGATTGAAT | 697del | InF | nd | nd | 1 | 2 (2) | |
| Cd1.18 | 664_668delinsTG | 690_697delinsGGAGCCTGCCGGAGCAA | InF | nd | nd | 2 | 1 (1) | |
| Cd1.19 | 666_671del | 693_697delinsTGCTCTGCAA | ps | nd | nd | 2 | 1 (3) | |
| Cd1.20 | 664_668delinsG | 675_694del | InF | nd | nd | 3 | 1 (4) | |
| Cd1.21 | 664_668delinsTG | 675_694del | ps | nd | nd | 3 | 1 (2) | |
| Cd1.22 | 666_697delinsGACGCAAGGACGCAA | * | ps | nd | nd | 2 | 1 (1) | |
| Cd2.1 | 665_666insGAGT | 689_695del | InF | nd | nd | 1, 1, 1 | 1 (2), 1 (5), 1 (1) | |
| Cd2.2 | 660_666del | 680_694del | InF | nd | nd | 1 | 1 (5) | |
| Cd2.3 | 664_665del | 691_693delinsAGGCTCTGCCGGCTCT | ps | nd | nd | 1 | 1 (2) | |
| LhCn | ||||||||
| Cn1.1 | 220_221insCGTCCT | 245_246delinsGTGGATGA | ps | 0, 2 | 1 (5), 1 (5) | 2 | 1 (2) | |
| Cn1.2 | 220_222del | 246_247insGAACACAAACACG | ps | 2, 3 | 4 (16), 3 (18) | 2, 3 | 4 (16), 2 (9) | |
| Cn1.3 | 220_221del | 244_246delinsG | ps | 0 | 1 (1) | nd | nd | |
| Cn1.4 | 221_223del | 244_247delinsG | InF | 0 | 1 (1) | nd | nd | |
| Cn1.5 | 223 T > A | 246_247insGAACATAACGGGAACATAAT | ps | 0 | 1 (2) | nd | nd | |
| Cn1.6 | 223_224del | 246del | InF | 0 | 1 (2) | nd | nd | |
| Cn2.1 | 219_227delinsATGTTTTC | 230_342del | InF | nd | nd | 1 | 1 (3) | |
| Cn2.2 | 182_228delinsAA | 234_246delinsACATCTCCTCTCTCTCA | InF | nd | nd | 1, 1 | 1 (1), 1 (5) | |
| Cn2.3 | 221_222delinsCA | 243_248del | InF | nd | nd | 1 | 1 (4) | |
| Cn2.4 | 221_222del | 243_248delinsAA | ps | nd | nd | 1 | 1 (1) | |
| Cn2.5 | 220_222del | 245_247delinsTGAACACAAACACG | ps | nd | nd | 1 | 1 (1) | |
| Cn2.6 | 220_222del | 246del | ps | nd | nd | 1 | 1 (5) | |
| Cn2.7 | 221_222insT | 247_248insTCGA | ps | nd | nd | 1 | 1 (5) | |
| Cn2.8 | 219del | 246_251delinsGCGGGAGCGGG | ps | nd | nd | 1 | 1 (1) | |
aMutation name defines the combination of alleles at sgRNA1 and sgRNA2 target sites in an individual as a single mutation.
bMutation nomenclature, showing the nucleic acid sequence changes in LhCd and LhCn, are based on the recommendations by the Human Genome Variation Society (http://www.hgvs.org/), with modification to omit the “prefix.” Mutations within 10 nucleotides were considered one mutation event.
cMutations for sgRNA1 corresponding to either LhCd or LhCn.
dMutations for sgRNA2 corresponding to either LhCd or LhCn.
eType of mutation (ps, premature stop; InF, in-frame mutation, nd, mutation type cannot be determined because the putative splice site was affected).
fnd = not determined.
*gMutation corresponding to sgRNA 1 that also spans the sgRNA 2 target site.
hwt = wild-type allele.
iSequence corresponding to the delins was too long for the respective table column: 649_665delinsTAAATTGTACAATTTATTGGCCAACTATTTCTAAAGACGGTTATCATGACATAAATACCTAATTTGGGGTTTTTGTCTGTGGGATATGCCTTACAGACTGAAAATCTATTGTTCCATTTCTCTTCTTC.