Table 2:
Open in new tab

Overview of ovoD-assisted gene targeting efficiency and precision of the unc-13 locus without and with the use of modified sgRNA sites

AlleleWith unmodified sgRNA sites in HDR plasmid, n/N (%)With modified sgRNA sites in HDR plasmid, n/N (%)
unc-13#2
 No. of clonal F1 offspring analyzed2120
  With edited missense mutation3/21 (14)11/20 (55)
  With correct unmodified/modified 5′-gRNA site sequence1/3 (33)11/11 (100)
  With correct unmodified/modified 3′-gRNA site sequence2/3 (67)10/11 (91)
unc-13#3
 No. of clonal F1 offspring analyzed1420
  With edited missense mutation3/14 (21)10/20 (50)
  With correct unmodified/modified 5′-gRNA site sequence2/3 (67)10/10 (100)
  With correct unmodified/modified 3′-gRNA site sequence0/3 (0)10/10 (100)
unc-13#4
 No. of clonal F1 offspring analyzed14Not applicable
  With edited missense mutation3/14 (21)
  With correct unmodified/modified 5′-gRNA site sequence0/3 (0)
  With correct unmodified/modified 3′-gRNA site sequence1/3 (33)
unc-13#5
 No. of clonal F1 offspring analyzed24Not applicable
  With edited missense mutation12/24 (50)
  With correct unmodified/modified 5′-gRNA site sequence0/8 (0)
  With correct unmodified/modified 3′-gRNA site sequence0/8 (0)
unc-13#6
 No. of clonal F1 offspring analyzed8Not applicable
  With edited missense mutation5/8 (63)
  With correct unmodified/modified 5′-gRNA site sequence0/4 (0)
  With correct unmodified/modified 3′-gRNA site sequence0/4 (0)
unc-13#7
 No. of clonal F1 offspring analyzed48Not applicable
  With edited missense mutation8/48 (17)
  With correct unmodified/modified 5′-gRNA site sequence2/4 (50)
  With correct unmodified/modified 3′-gRNA site sequence0/4 (0)
AlleleWith unmodified sgRNA sites in HDR plasmid, n/N (%)With modified sgRNA sites in HDR plasmid, n/N (%)
unc-13#2
 No. of clonal F1 offspring analyzed2120
  With edited missense mutation3/21 (14)11/20 (55)
  With correct unmodified/modified 5′-gRNA site sequence1/3 (33)11/11 (100)
  With correct unmodified/modified 3′-gRNA site sequence2/3 (67)10/11 (91)
unc-13#3
 No. of clonal F1 offspring analyzed1420
  With edited missense mutation3/14 (21)10/20 (50)
  With correct unmodified/modified 5′-gRNA site sequence2/3 (67)10/10 (100)
  With correct unmodified/modified 3′-gRNA site sequence0/3 (0)10/10 (100)
unc-13#4
 No. of clonal F1 offspring analyzed14Not applicable
  With edited missense mutation3/14 (21)
  With correct unmodified/modified 5′-gRNA site sequence0/3 (0)
  With correct unmodified/modified 3′-gRNA site sequence1/3 (33)
unc-13#5
 No. of clonal F1 offspring analyzed24Not applicable
  With edited missense mutation12/24 (50)
  With correct unmodified/modified 5′-gRNA site sequence0/8 (0)
  With correct unmodified/modified 3′-gRNA site sequence0/8 (0)
unc-13#6
 No. of clonal F1 offspring analyzed8Not applicable
  With edited missense mutation5/8 (63)
  With correct unmodified/modified 5′-gRNA site sequence0/4 (0)
  With correct unmodified/modified 3′-gRNA site sequence0/4 (0)
unc-13#7
 No. of clonal F1 offspring analyzed48Not applicable
  With edited missense mutation8/48 (17)
  With correct unmodified/modified 5′-gRNA site sequence2/4 (50)
  With correct unmodified/modified 3′-gRNA site sequence0/4 (0)

Notes: For experimental sets using unmodified sgRNA sites, “correct” refers to their wild-type sequence, and for experiments using modified sgRNA sites, “correct” refers to the modified sequence.

Table 2:
Open in new tab

Overview of ovoD-assisted gene targeting efficiency and precision of the unc-13 locus without and with the use of modified sgRNA sites

AlleleWith unmodified sgRNA sites in HDR plasmid, n/N (%)With modified sgRNA sites in HDR plasmid, n/N (%)
unc-13#2
 No. of clonal F1 offspring analyzed2120
  With edited missense mutation3/21 (14)11/20 (55)
  With correct unmodified/modified 5′-gRNA site sequence1/3 (33)11/11 (100)
  With correct unmodified/modified 3′-gRNA site sequence2/3 (67)10/11 (91)
unc-13#3
 No. of clonal F1 offspring analyzed1420
  With edited missense mutation3/14 (21)10/20 (50)
  With correct unmodified/modified 5′-gRNA site sequence2/3 (67)10/10 (100)
  With correct unmodified/modified 3′-gRNA site sequence0/3 (0)10/10 (100)
unc-13#4
 No. of clonal F1 offspring analyzed14Not applicable
  With edited missense mutation3/14 (21)
  With correct unmodified/modified 5′-gRNA site sequence0/3 (0)
  With correct unmodified/modified 3′-gRNA site sequence1/3 (33)
unc-13#5
 No. of clonal F1 offspring analyzed24Not applicable
  With edited missense mutation12/24 (50)
  With correct unmodified/modified 5′-gRNA site sequence0/8 (0)
  With correct unmodified/modified 3′-gRNA site sequence0/8 (0)
unc-13#6
 No. of clonal F1 offspring analyzed8Not applicable
  With edited missense mutation5/8 (63)
  With correct unmodified/modified 5′-gRNA site sequence0/4 (0)
  With correct unmodified/modified 3′-gRNA site sequence0/4 (0)
unc-13#7
 No. of clonal F1 offspring analyzed48Not applicable
  With edited missense mutation8/48 (17)
  With correct unmodified/modified 5′-gRNA site sequence2/4 (50)
  With correct unmodified/modified 3′-gRNA site sequence0/4 (0)
AlleleWith unmodified sgRNA sites in HDR plasmid, n/N (%)With modified sgRNA sites in HDR plasmid, n/N (%)
unc-13#2
 No. of clonal F1 offspring analyzed2120
  With edited missense mutation3/21 (14)11/20 (55)
  With correct unmodified/modified 5′-gRNA site sequence1/3 (33)11/11 (100)
  With correct unmodified/modified 3′-gRNA site sequence2/3 (67)10/11 (91)
unc-13#3
 No. of clonal F1 offspring analyzed1420
  With edited missense mutation3/14 (21)10/20 (50)
  With correct unmodified/modified 5′-gRNA site sequence2/3 (67)10/10 (100)
  With correct unmodified/modified 3′-gRNA site sequence0/3 (0)10/10 (100)
unc-13#4
 No. of clonal F1 offspring analyzed14Not applicable
  With edited missense mutation3/14 (21)
  With correct unmodified/modified 5′-gRNA site sequence0/3 (0)
  With correct unmodified/modified 3′-gRNA site sequence1/3 (33)
unc-13#5
 No. of clonal F1 offspring analyzed24Not applicable
  With edited missense mutation12/24 (50)
  With correct unmodified/modified 5′-gRNA site sequence0/8 (0)
  With correct unmodified/modified 3′-gRNA site sequence0/8 (0)
unc-13#6
 No. of clonal F1 offspring analyzed8Not applicable
  With edited missense mutation5/8 (63)
  With correct unmodified/modified 5′-gRNA site sequence0/4 (0)
  With correct unmodified/modified 3′-gRNA site sequence0/4 (0)
unc-13#7
 No. of clonal F1 offspring analyzed48Not applicable
  With edited missense mutation8/48 (17)
  With correct unmodified/modified 5′-gRNA site sequence2/4 (50)
  With correct unmodified/modified 3′-gRNA site sequence0/4 (0)

Notes: For experimental sets using unmodified sgRNA sites, “correct” refers to their wild-type sequence, and for experiments using modified sgRNA sites, “correct” refers to the modified sequence.

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