| ZKO number |
ZKO614 |
| Recent progress of this line |
F2 generation has been raised
|
|
Gene name |
sts
|
|
Ensembl ID |
ENSDARG00000053241
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GGGATATCTGCCTGTGAGAAagg
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
Exon 3
|
|
Restriction enzyme (RE) |
EcoR V (GAT/ATC)
|
|
PCR primer (forward) |
5’- ATGGCAACCACTAAAAGCAGAA -3’
|
|
PCR primer (reverse) |
5’- CTCTGGAAGGGGTGAAACTGAC -3’
|
|
Length of expected PCR product (bp)
|
444
|
|
Length of expected RE digestion product (bp) |
360 bp +84 bp
|
|
Plasmids used for making Cas9 mRNA |
pXT7-hCAS9
|
| Plasmids?used?for?making?gRNA |
pMD19T-gRNA
|
| Primers?used?for?making?gRNA |
Pr-ZKO 614
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
300
|
|
Dosage of injected gRNA (ng/ul)
|
100
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
76
|
|
Method to estimate mutation efficiency |
sequencing of PCR product
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO614.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
20
|
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture) |
|
|
Mutation type
|
Indel
|
|
Germline transmission efficiency in adult F0
|
4
|
|
Wild-type (WT) sequence
|
CTGATCGGATGGGATATCTGCCTGTGAGAAAGG
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
CTGCTTGTxxxxxxxxxxxxxxxxTCAGAAAGG (+3 bp, -19 bp)
|
|
F0-2
|
GGGxxxxxxxxxxxxxxxxxxxx( -20 bp)
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
caaatct---------------------------------------------------------CCCTGCTGGGAGT (-57 bp)
|
|
F0-2
|
aacaaatta--------cCTGATCGGATGGCCTGTGAGAAAGGCTGCCCTGCTGGGAGTGCAGG(-8bp)
|
|
Allele name and predicted product
|
614a
614b |
caaatct---------------------------------------------------------CCCTGCTGGGAGT in frame(delate aa 13)
|
| aacaaatta--------cCTGATCGGATGGCCTGTGAGAAAGGCTGCCCTGCTGGGAGTGCAGG Truncated protein (136aa) |
|
突变创建时间 |
2013-04-14
|
|
填表时间 |
2013/12/23
|
|
修改时间 |
2016/10/24
|
|
PI/负责人 (工作单位) |
|
