| ZKO number |
ZKO972 |
| Recent progress of this line |
F1 progeny has been raised
|
|
Gene name |
tbck
|
|
Ensembl ID |
ENSDARG00000013667
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
antisense strand
5’- GGAAGTCGACATCTGCTCCA tgg -3’
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
Exon 5
|
|
Restriction enzyme (RE) |
T7EI
|
|
PCR primer (forward) |
5’- ATAAACTAACTTCAAACCCAAC -3’
|
|
PCR primer (reverse) |
5’- GACGATGCACATCTAATGTTAG -3’
|
|
Length of expected PCR product (bp)
|
453
|
|
Length of expected RE digestion product (bp) |
300,150
|
|
Plasmids used for making Cas9 mRNA |
pXT7-hCAS9
|
| Plasmids used for making gRNA |
pMD19T-gRNA
|
| Primers used for making gRNA |
Pr-ZKO 972
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
300
|
|
Dosage of injected gRNA (ng/ul)
|
60
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
80
|
|
Method to estimate mutation efficiency |
T7EI detection
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO972.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
|
15%
|
|
Wild-type (WT) sequence
|
5'- TATGACCGACCATGGAGCAGATGTCGACTTCCCAATT -3'
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
|
|
F0-2
|
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
5'-TATGACCGA--------CAGATGTCGACTTCCCAATT -3' (-8 bp)
|
|
F0-2
|
5'- ATATGACCGACCATGGtcatatggccGAGCAGATGTC -3' (+10 bp)
|
|
Allele name and predicted product
|
972a
972b |
5'- TATGACCGA--------CAGATGTCGACTTCCCAATT -3' (-8 bp) Truncated protein (172aa)
|
| 5'- ATATGACCGACCATGGtcatatggccGAGCAGATGTC -3' (+10 bp) Truncated protein 178 aa |
|
突变创建时间 |
|
|
填表时间 |
2013/12/24
|
|
修改时间 |
2016/8/8
|
|
PI/负责人 (工作单位) |
|