Table 4 Segregation patterns of CRISPR/Cas9-induced mutations in the T1generation.

From: CRISPR/Cas9-mediated genome editing efficiently creates specific mutations at multiple loci using one sgRNA in Brassica napus

Target gene	sgRNA	Line	T0			T1
Target gene	sgRNA	Line	Zygosity^◐	Genotype	Cas9^§	Segregation ratio	P value	Cas9^§
BnaA6.RGA	BnaRGA-sgRNA1	L40	Homozygote	d1d1	+	8 d1d1		7(+), 1(−)
BnaA6.RGA	BnaRGA-sgRNA1	L44	Homozygote	i1i1	+	17 i1i1		12(+), 5(−)
BnaA9.RGA	BnaRGA-sgRNA1	L44	Homozygote	d6d6	+	17 d6d6		12(+), 5(−)
BnaA6.RGA	BnaA6.RGA-sgRNA1	L5	Bi-allele	d5, d6	+	9 d5d5: 9 d5d6:2 d6d6^※	0.08*	16(+), 4(−)
BnaA6.RGA	BnaA6.RGA-sgRNA1	L6	Bi-allele	d5, d9	+	4 d5d5: 5 d5d9:0 d9d9^※	0.02*	7(+), 2(−)
BnaA6.RGA	BnaRGA-sgRNA1	L46	Bi-allele	i1, d5	+	6 i1i1: 9 i1d5: 2 d5d5	0.38	15(+), 2(−)
BnaA9.RGA	BnaRGA-sgRNA1	L27	Bi-allele	d1, d5	+	0 d1d1:20 d1d5:0 d5d5	3.1E-07*	16(+), 4(−)
BnaC7.RGA	BnaRGA-sgRNA1	L38	Bi-allele	i1, d2	+	3 i1i1: 8 i1d2: 4 d2d2	0.90	12(+), 3(−)
BnaC5.DA1	BnaC5.DA1-sgRNA1	L1	Bi-allele	i1a, i1b	+	11 i1ai1a:1 i1ai1b: 2 i1bi1b	2.3E-07*	14(+)
BnaC5.DA1	BnaC5.DA1-sgRNA1	L16	Bi-allele	i1a, i1b	−	4 i1ai1a: 8 i1ai1b: 4 i1bi1b	1.0	16(−)
BnaA2.DA2	BnaDA2-sgRNA2	L12	Bi-allele	i1, d4	+	0 i1i1: 2 i1d4: 13 d4d4	0.04*	15(+)
BnaC6.DA2	BnaDA2-sgRNA2	L20	Bi-allele	i1a, i1b	+	3 i1ai1a:12 i1ai1b: 0 i1bi1b	1.8E-05*	13(+), 2(−)
BnaA9.FUL	BnaFUL-sgRNA2	L15	Bi-allele	i1, d1	+	5 i1i1: 10 i1d1: 1 d1d1	0.008*	13(+), 3(−)
BnaC7.FUL	BnaFUL-sgRNA2	L23	Bi-allele	i2, d5	+	3 i2i2: 8 i2d5: 5 d5d5	0.77	15(+), 1(−)
BnaA6.RGA	BnaRGA-sgRNA1	L33	Heterozygote	i1, WT	+	3 i1ai1a: 7 i1ai1b: 2 i1bi1b: 1 i1aWT	0.31	12(+), 1(−)
BnaA9.RGA	BnaRGA-sgRNA1	L40	Heterozygote	d22, WT	+	7 i1i1: 2 d22d22: 7 h^※		15(+), 1(−)
BnaC7.RGA	BnaRGA-sgRNA1	L40	Heterozygote	i1a, WT	+	5 i1ai1a: 9 i1ai1b: 2 i1bi1b	0.31	15(+), 1(−)
BnaA6.DA1	BnaA6.DA1-sgRNA2	L6	Heterozygote	i1, WT	+	3 i1i1: 10 i1WT: 2 WTWT	0.90	15(+)
BnaA6.DA1	BnaA6.DA1-sgRNA2	L24	Heterozygote	d1, WT	+	4 d1d1: 6 d1WT: 3 WTWT		13(+)
BnaA2.DA2	BnaDA2-sgRNA2	L1	Heterozygote	i1, WT	+	11 i1ai1b: 5 WTWT	0.06*	12(+), 4(−)
BnaA2.DA2	BnaDA2-sgRNA2	L20	Heterozygote	i1, WT	+	3 i1ai1a: 6 i1aWT: 4 i1ai1b: 2 WTWT		13(+), 2(−)
BnaC6.DA2	BnaDA2-sgRNA2	L1	Heterozygote	i1, WT	+	0 i1i1: 10 i1WT: 6 WTWT	0.31	12(+), 4(−)
BnaA6.RGA	BnaA6.RGA-sgRNA2	L4	Chimera	d6, d5, WT	+	3d5d5: 16 h^※		15(+), 4(−)
BnaC9.RGA	BnaRGA-sgRNA1	L27	Chimera	d1, d5, d33, d36	+	2d1 h:10 d5 h: 2d33 h: 5d36 h^※		15(+), 4(−)
BnaC9.RGA	BnaRGA-sgRNA1	L40	Chimera	i1, d5, WT	+	16 d19h^※		15(+), 1(−)
BnaC6.DA2	BnaDA2-sgRNA1	L12	Chimera	d4, h	+	5 d4d4: 2 d4h: 2 d7h: 3 d3d4: 1 WTWT: 2 h^※		15(+)
BnaA9.RGA	BnaRGA-sgRNA1	L43	WT	WT^♢	+	10 WT WT		10(+)
BnaA6.DA1	BnaA6.DA1-sgRNA2	L15	WT	WT^♢	+	10 WT WT		10(+)
BnaA6.DA1	BnaA6.DA1-sgRNA2	L25	WT	WT^♢	+	10 WT WT		10(+)

◐The zygosity of the homozygote, bi-allele, and heterozygote in T0 plants. ^♢WT, no mutations were identified. ^§Presence of Cas9 sequence: +, Cas9 positive; −, Cas9 negative; n.d., Not determined. ^※More data are needed to fully explain the T1 genotypes. d#, # of bp deleted from a target site; d#a, same number of deletion at one site; d#b, same number of deletion at other sites; i#, # of bp inserted at target site; i#a, same number of insertion at one site; i#b, same number of insertion of different nucleotide at the same site; c#, combined mutation; h, heterogeneous, more than one sequence detected in the sample. χ² test *P value < 0.1.

Back to article page

Table 4 Segregation patterns of CRISPR/Cas9-induced mutations in the T1generation.

Search

Quick links