| ZKO number |
ZKO1141 |
| Recent progress of this line |
F2 individuals have been indentified
|
|
Gene name |
samd1b
|
|
Ensembl ID |
ENSDARG00000079062
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
TCTACCCGCGAGGGGGATGGggg
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
exon1
|
|
Restriction enzyme (RE) |
n/a
|
|
PCR primer (forward) |
AATGGATGCTGTCCGAAGTA
|
|
PCR primer (reverse) |
AAGAGCGGAAATCAGAAGGT
|
|
Length of expected PCR product (bp)
|
449
|
|
Length of expected RE digestion product (bp) |
|
|
Plasmids used for making Cas9 mRNA |
pGH-T7-zCAS9
|
| Plasmids used for making gRNA |
pT7-gRNA-ZKOpT7-gRNA-ZKO
|
| Primers used for making gRNA |
Pr-ZKO 1141
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
650
|
|
Dosage of injected gRNA (ng/ul)
|
69
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
90
|
|
Method to estimate mutation efficiency |
sequencing of PCR product
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO1141.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
72
|
Picture from F0 embryos
(gel picture or sequencing picture) |
|
|
Mutation type
|
deletion
|
|
Germline transmission efficiency in adult F0
|
50%
|
|
Wild-type (WT) sequence
|
TCGGCCACATCACTCTACCCGCGAGGGGGATGGGGGCGGCCAAGCC
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
TCGGCCACATCACTCTACCCGCGAGGGGGA-------GGCCAAGCC(-7bp)
|
|
F0-2
|
TCGGCCACATCACTCTACCCGCGAGGGG-----------CCAAGCC(-11bp)
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
TCGGCCACATCACTCTACCCGCGAGGGGGA-------GGCCAAGCC(-7bp)
|
|
F0-2
|
TCGGCCACATCACTCTACCCGCGAGGGG-----------CCAAGCC(-11bp)
|
|
Allele name and predicted product
|
1141a
1141b |
AGCGGATCAC--------AGAGCTGAAGGATGA ; Truncated protein 396aa
|
| AGCGGATCAC-----AGCTTTCTGCTGA; no stop coden |
|
突变创建时间 |
2014-07-31
|
|
填表时间 |
2013/12/20
|
|
修改时间 |
2016/8/9
|
|
PI/负责人 (工作单位) |
|