Table 3 Effect of target sequence on the efficiency of targeted mutagenesis of the GFP transgene.

From: CRISPR/Cas9-mediated targeted mutagenesis in Japanese cedar (Cryptomeria japonica D. Don)

Target∗

No. of transformed lines (A)

No. of GFP-knock-out lines∗∗ (B)

Mutation frequency (%)¦

No. of lines having multiple mutation patterns§ (C)

Mosaicism frequency (%)

#1

136¿

34¿

25.0¿,b

15

44.1c

#2_rev

70

29

41.4a

17

58.6c

#3_rev

77

24

31.2a,b

15

62.5c

  1. ET embryogenic tissue.
  2. ∗pZK_FFCas9 vectors with gRNA expression cassettes driven by AtU6-26 promoter were transformed into ET.
  3. ∗∗The number of ET lines was counted after 87 days of selective cultivation.
  4. ¦(B/A) × 100. Pairwise multiple comparison of proportions was performed using Tukey’s multiple comparison test. Proportions with significant difference were labeled with different letters (P < 0.05).
  5. §The number of ET lines having multiple mutation patterns was counted.
  6. (C/B) × 100. Pairwise multiple comparison of proportions was performed using Tukey’s multiple comparison test. Proportions with a significant difference were labeled with different letters (P < 0.05).
  7. ¿Data from Table 1.
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