Table 2 cku-80 RNAi results in increased knock-in efficiency
P0 strainpha-1 oligoRepair oligoViable injected P0pha-1 rescued F1P0 with rescued F1PCR hitsKnock-insKnock-ins/F1 rescue (%)Knock-ins/P0 (%)
pha-1(ts); control(RNAi)a80mernhr-23:::2×FLAG101111100.010.0
pha-1(ts); cku-80(RNAi)a80mernhr-23:::2×FLAG161066550.031.3
pha-1(ts); control(RNAi)b200mernhr-25:::2×FLAG2174000.00.0
pha-1(ts); cku-80(RNAi)b200mernhr-25:::2×FLAG2236125411.118.2
pha-1(ts); control(RNAi)b200mernhr-23::3xFLAG34741114.33.0
pha-1(ts); cku-80(RNAi)b200mernhr-23::3xFLAG13532240.015.4
pha-1(ts); cku-80(RNAi)b200mer2×FLAG::smo-115297141138.073.3
lig-4 stop000.00.0
  • Summary of pha-1(ts) coselection experiments testing NHEJ inactivation. The RNAi treatment of the injected P0 animals is indicated. Viable P0 are the number of injected animals that produced eggs; a variable number of animals are sterile in each experiment. The length of the pha-1(ts) sense repair oligo is provided. For nhr-23::2×FLAG experiments, animals were injected with 60 ng/µl each of the pha-1 and nhr-23 CRISPR/Cas9 plasmids, and 50 ng/µl each of the pha-1(ts) repair and nhr-23::2×FLAG (sense) oligos. For the remaining experiments, animals were injected with 50 ng/µl of the pha-1 CRISPR/Cas9 plasmid, 25 ng/µl of appropriate PU6::sgRNA template PCR product, and 50 ng/µl each of the pha-1(ts) repair oligo and knock-in oligo.

  • a Pre-RNAi diet = OP50 E. coli.

  • b Pre-RNAi diet = HB101 E. coli.