| ZKO number |
ZKO1156 |
| Recent progress of this line |
F2 individuals have been indentified
|
|
Gene name |
C3 (7 of 9)
|
|
Ensembl ID |
C3 (7 of 9) ENSDARG00000090176
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GGATTTCGAGAGCGCCGGGC
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
exon4
|
|
Restriction enzyme (RE) |
n/a
|
|
PCR primer (forward) |
5’- TCTGTTAAACATCATTCGGGA -3’
|
|
PCR primer (reverse) |
5’- ATGCACATCCATCATCAGCA -3’
|
|
Length of expected PCR product (bp)
|
473
|
|
Length of expected RE digestion product (bp) |
|
|
Plasmids used for making Cas9 mRNA |
pSP6-2sNLS-spCas9
|
| Plasmids used for making gRNA |
pT7-gRNA
|
| Primers used for making gRNA |
Pr-ZKO
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
563
|
|
Dosage of injected gRNA (ng/ul)
|
127.6
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
90
|
|
Method to estimate mutation efficiency |
sequencing of PCR product
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO1156.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
81.8
|
Picture from F0 embryos
(gel picture or sequencing picture) |
|
|
Mutation type
|
Indel
|
|
Germline transmission efficiency in adult F0
|
100% (2/2: Number of positive F0 / Total number of F0 screened)
|
|
Wild-type (WT) sequence
|
tggcagctaaGGATTTCGAGAGCGCCGGGCaggtggtgcgctg
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
tggcagctaaGGATTTCGAGAGCGCxxxxxaggtggtgcgctg (-5 bp)
|
|
F0-2
|
tggcagctaaGGATTTCGAGAGCctcatcactcgtttaaGGGCaggtggtgcgctg (+16 bp, -3 bp)
tggcagctaaGGATTTCGAGxxxxxxxxGCaggtggtgcgctg (-8 bp)
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
33.3% (1/3: Number of F1 carrier / Total number of F1 screened)
tggcagctaaGGATTTCGAGAGCGCxxxxxaggtggtgcgctg (-5 bp)
|
|
F0-2
|
66.7% (2/3: Number of F1 carrier / Total number of F1 screened)
tggcagctaaGGATTTCGAGxxxxxxxxGCaggtggtgcgctg (-8 bp)
tggcagctaaGGATTTCGAGAGCGCCGxxxxxxtggtgcTctg (-6 bp)
|
|
Allele name and predicted product
|
1156a
1156b |
tggcagctaaGGATTTCGAGAGCGCxxxxxaggtggtgcgctg; Truncated protein (190 aa)
|
| tggcagctaaGGATTTCGAGxxxxxxxxGCaggtggtgcgctg; Truncated protein (189 aa) |
|
突变创建时间 |
2014-08-01
|
|
填表时间 |
2013/12/20
|
|
修改时间 |
2016/8/9
|
|
PI/负责人 (工作单位) |
|