ZKO number |
ZKO1130 |
Recent progress of this line |
F2 generation has been raised
|
Gene name |
null
|
Ensembl ID |
ENSDARG00000089993
|
Zebrafish strain |
AB
|
Cas9 target site 1 sequence |
sense strand
GGAAAATGGACGGGGATTCC
|
Cas9 target site 2 sequence |
|
Position in the gene |
Exon 3
|
Restriction enzyme (RE) |
n/a
|
PCR primer (forward) |
5’-GATCAGTCTACTGGACCAAACATGC-3’
|
PCR primer (reverse) |
5’-TTTCACATTTCCTGACTGGTTTTTG-3’
|
Length of expected PCR product (bp)
|
527
|
Length of expected RE digestion product (bp) |
|
Plasmids used for making Cas9 mRNA |
pXT7-hCAS9
|
Plasmids?used?for?making?gRNA |
pMD19T-gRNA
|
Primers?used?for?making?gRNA |
Pr-ZKO 1130
|
Dosage of injected Cas9 mRNA (ng/ul)
|
100
|
Dosage of injected gRNA (ng/ul)
|
50
|
Survival rate of F0 embryo @ 1dpf (%)
|
44
|
Method to estimate mutation efficiency |
sequencing of PCR product
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO1130.pdf
|
Mutation efficiency in F0 embryos (%)
|
50
|
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture) |
|
Mutation type
|
deletion
|
Germline transmission efficiency in adult F0
|
29% (2/7)
|
Wild-type (WT) sequence
|
GGTGGAAAATGGACGGGGATTCCTGGAGTT
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
|
F0-2
|
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
GGTGGAAAATGGACGGGGA--------GTT(-8,+0)
|
F0-2
|
GGTGGAAAATGGACGGGGATTCAGCTGGAGTT(-0,+2)
|
Allele name and predicted product
|
1130a 1130b |
GGTGGAAAATGGACGGGGA--------GTT Truncated protein (112aa)
|
GGTGGAAAATGGACGGGGATTCAGCTGGAGTT Truncated protein (129 aa) |
突变创建时间 |
2013-11-10
|
填表时间 |
2013/12/20
|
修改时间 |
2016/8/11
|
PI/负责人 (工作单位) |
|