| ZKO number |
ZKO1149 |
| Recent progress of this line |
F2 generation has been raised
|
|
Gene name |
asf1bb
|
|
Ensembl ID |
asf1bb ENSDARG00000043713
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GGTGCTGAATGTGGCTGTTC
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
exon1
|
|
Restriction enzyme (RE) |
n/a
|
|
PCR primer (forward) |
5’- GCTCCACTCGAGGATCGTTT -3’
|
|
PCR primer (reverse) |
5’- CGCACCTACAGTCTTGGGAA -3’
|
|
Length of expected PCR product (bp)
|
360
|
|
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)
|
50
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
80
|
|
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/ZKO1149.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
28
|
Picture from F0 embryos
(gel picture or sequencing picture) |
|
|
Mutation type
|
deletion
|
|
Germline transmission efficiency in adult F0
|
20%
|
|
Wild-type (WT) sequence
|
GTGCAGGTGCTGAATGTGGCTGTTCTGGATAACCC
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
TAAAGTGCAGGTGCTGAATGTGG---------ATAACCCCA（-9bp deletion）
|
|
F0-2
|
GTGCAGGTGCTGAATGTGGCTG-----GATAACCCCAG（5bp deletion）
GTGCAGGTGCTGAATGTGGCTGGATAACCTGGATAACCC
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
GTGCAGGTGCTGAAT--------TCTGGATAACCC(-8bp deletion)
|
|
F0-2
|
GTGCAGGTGCTGAATGTGGCTG----GGATAACCC(-4bp deletion)
|
|
Allele name and predicted product
|
1149a
1149b |
AAAGTGCAGGTGCTGAAT--------TCTGGATAACCC(-8bp)Truncated protein
|
| AAAGTGCAGGTGCTGAATGTGGCTG----GGATAACCCCA(-4bp)truncated protein |
|
突变创建时间 |
2014-06-20
|
|
填表时间 |
2013/12/20
|
|
修改时间 |
2016/8/9
|
|
PI/负责人 (工作单位) |
|