Table 1 Summary of genome-editing experiments across nematode species, using TALENs and CRISPR/Cas9
TargetDSB-inducing nucleaseSpeciesDesired changeKnock-in sequenceAnimals injected (1 gonad arm)F1sAll mutations (% of F1s)Knockouts (%)Total knock-ins (%)Precise knock-ins (% of all mutations)
ben-1TALENsC. elegansPsiI → NcoICCATGG1554822 (4.0)10126 (27)
ben-1TALENsC. elegansPsiI → NcoICCATGG1044012 (2.7)66 (1.4)2 (17)
PsiI → EcoRIGAATTC3 (25)
PsiI → HindIIIAAGCTT1 (8)
ben-1TALENsC. elegans+ MEXAAGCGCAGGAG123827 (1.8)344 (57)
smo-1TALENsC. elegansL88KAAG185161 (0.2)011 (100)
smo-1TALENsC. elegansL88KAAAGCGCT147292 (0.3)022 (100)
rex-1TALENsC. elegansKnockout, NHEJ1247912 (2.5)12 (2.5)
rex-1TALENsC. elegansKnockout, Δ77 bp, homology-directed repairCAT1564885 (13.1)612412 (14)
NHEJHDR-mediated Δ by insertion
rex-32TALENsC. elegans+ 3′-FRTSame as 5′-FRT1284712 (1.4)399 (75)
rex-32FLP-FRTC. elegansKnockout by FLP186114 (0.7)4 (0.7)
sdc-2TALENsC. sp. 9 (JU1422)Knockout, NHEJ42421 (2.4)1 (2.4)
unc-119TALENsP. pacificus
(PS312)Knockout, NHEJ1650134 (6.8)34 (6.8)
unc-119TALENsP. pacificus+ HindIIIAAGCTT1132718 (5.5)1263 (17)
unc-119TALENsP. pacificus+ HA tagTACCCATACGACGTCCCAGATTACGCT1550412 (2.3)755 (42)
gfpCas9C. elegansKnockout, NHEJ216868 (1.2)8 (1.2)
  • Shown are data from all the genome-editing experiments described in the text. The target locus, the DSB-inducing agent, the species, and the types of modifications, including their quantification, are provided. For the knock-in experiments, the inserted sequence is listed, as indicated by “+”. For all experiments, the number of animals injected is provided along with the number of F1s screened from the 8- to 16-hr window postinjection. Because only one of the two gonad arms was injected in each animal, only half the F1s scored had the possibility of receiving a mutation. The frequency was not corrected for this fact. For both knock-in and knockout experiments, the total number of mutations recovered is shown, along with the percentage of total mutations relative to number of F1s screened. For knock-in experiments, three classes of mutations were found: NHEJ-mediated deletions, HDR-mediated imprecise insertions, and HDR-mediated precise insertions. Shown are the total number of knock-ins (precise and imprecise), the number of precise knock-ins, and the number of NHEJ-mediated deletions. The percentage of precise knock-in mutations relative to the total number of mutations is also shown.