Table 1 Heritability of dominant mutant phenotypes in F1 animals and their cosegregation with a fluorescent transformation marker
sgRNA and Recombination TemplateNumber of Mutant F1sHeritable (%)Of Heritable, Fluorescent (%)
Oligonucleotide templates
 sqt-1 sgRNA #1 sense6862 (91)2 (4)a
 sqt-1 sgRNA #1 antisense22 (100)0
 sqt-1 sgRNA #2 sense2519 (76)0
 sqt-1 sgRNA #2 antisense31 (33)0
 sqt-1 sgRNA #3 sense3631 (86)4 (13)
 sqt-1 sgRNA #3 antisense167 (44)1 (14)
 lin-12 sense12542 (34)12 (29)
 lin-12 antisense734 (5)3 (75)
 dpy-10 sense273143 (52)53 (37)
 dpy-10 antisense198118 (60)36 (31)
 Total oligonucleotide819429 (52)111 (27)a
Double-stranded circular templates
 sqt-1 sgRNA #2999 (9)0
 lin-1212535 (28)14 (40)
 Total double-stranded DNA22444 (20)14 (32)
Single-stranded circular templates
 sqt-1 sgRNA #210515 (14)0
 lin-1211827 (23)9 (33)
 Total single-stranded DNA22342 (19)9 (21)
  • The table shows the total number of mutant (roller or multivulva) animals resulting from microinjections with three sgRNAs targeting sqt-1, one sgRNA targeting lin-12, one sgRNA targeting dpy-10, and the Cas9 driven by the ubiquitous promoter of the eft-3 gene. Each mix was injected into 20-30 P0 animals in triplicate, except for dpy-10 sgRNA containing mixes, which were injected into 10-15 P0 animals in triplicate. Heritable changes are those that mutant animals segregate in the F2 generation (roller, dumpy, multivulva, and egg-laying deficient, respectively). The last column shows the proportion of F1 animals with heritable mutations which were also positive for the transgenic array.

  • The “Total oligonucleotide” row includes results of 30 separate microinjections with sense or antisense oligonucleotides as recombination templates.

  • a Fluorescence status was known for just 412 animals. The “Total double-stranded DNA” and “Total single-stranded DNA” rows include results of 6 separate injections each.