| ZKO number |
ZKO1157 |
| Recent progress of this line |
F2 individuals have been indentified
|
|
Gene name |
si:dkey-76b14.5
|
|
Ensembl ID |
C3 (5 of 9) ENSDARG00000053462
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
antisense strand
GGGATGTCCTTCGATGTCTC
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
exon9
|
|
Restriction enzyme (RE) |
n/a
|
|
PCR primer (forward) |
5’- AGGCAGTGAAATGGTGGATG -3’
|
|
PCR primer (reverse) |
5’- TTTAGTGATGCCGTTGGGTT -3’
|
|
Length of expected PCR product (bp)
|
302
|
|
Length of expected RE digestion product (bp) |
|
|
Plasmids used for making Cas9 mRNA |
pGH-T7-zCAS9
|
| Plasmids used for making gRNA |
pMD19T-gRNA
|
| Primers used for making gRNA |
Pr-ZKO
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
600
|
|
Dosage of injected gRNA (ng/ul)
|
100
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
90
|
|
Method to estimate mutation efficiency |
PCR and single clone sequencing
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO1157.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
80
|
Picture from F0 embryos
(gel picture or sequencing picture) |
|
|
Mutation type
|
Indel
|
|
Germline transmission efficiency in adult F0
|
50%
|
|
Wild-type (WT) sequence
|
AACCCGGGATGTCCTTCGATGTCTCGGTAAAA
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
AA---------TCCTTCGATGTCTC (-9 bp)
|
|
F0-2
|
AAACCCGGGTCCTTCAAACTTCAGTCCTTCGATGTCTCGGTAAAA (+14bp)
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
CAAACCCGGCAATTCTTCATCCTTCGATGTCTCGGTAAAA(+ 6bp,3bp mutation)
|
|
F0-2
|
CAAACCCGGAGAATTCGTCCTTCGATGTCTCGGTAAA
(+4bp, 1bp mut)
|
|
Allele name and predicted product
|
1157a
1157b |
CAAACCCGGAGAATTCGTCCTTCGATGTCTCGGTAAA (+4bp,1bp mutant)truncated protein
|
| CAAACCCGGCAATTCTTCATCCTTCGATGTCTCGGTAAAA(+ 6bp,3bp mutation)truncated protein |
|
突变创建时间 |
2014-04-18
|
|
填表时间 |
2013/12/20
|
|
修改时间 |
2016/8/9
|
|
PI/负责人 (工作单位) |
|