| ZKO number |
ZKO54 |
| Recent progress of this line |
F2 individuals have been indentified
|
|
Gene name |
mir459
|
|
Ensembl ID |
ENSDARG00000080051
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GGGCTTTTCTGGGCTCTAGagg,GGACAGGATCGGCCCAGGAGagg
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
upstream and downstream to delete the mirRNA
|
|
Restriction enzyme (RE) |
n/a
|
|
PCR primer (forward) |
gcgtaaaacatatggtggaacagt
|
|
PCR primer (reverse) |
TCACCCTTGAGGGAGAGAAGT
|
|
Length of expected PCR product (bp)
|
3286
|
|
Length of expected RE digestion product (bp) |
no RE is used
|
|
Plasmids used for making Cas9 mRNA |
pGH-T7-zCAS9
|
| Plasmids?used?for?making?gRNA |
pMD19T-gRNA
|
| Primers?used?for?making?gRNA |
Pr-ZKO 54
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
500
|
|
Dosage of injected gRNA (ng/ul)
|
100
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
80
|
|
Method to estimate mutation efficiency |
sequencing of PCR product
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO54.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
15
|
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture) |
|
|
Mutation type
|
deletion
|
|
Germline transmission efficiency in adult F0
|
20%
|
|
Wild-type (WT) sequence
|
ccaactaaagtaactccagggcttttctgggctctagagg<2738bp>ggagaggctgagagatggggacccgtgttgtttacctgtcat
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
ccaactaaagtaactccagggcttttctgggggagaggctgagagatggggacccgtgttgtttacctgtcat（-2747bp）
|
|
F0-2
|
ccaactaaagtaactccagggcttttctgggctctagagggaggctgagagatggggacccgtgttgtttacctgtcat(-2741bp)
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
ccaactaaagtaactccagggcttttctgggggagaggctgagagatggggacccgtgttgtttacctgtcat（-2747bp）
|
|
F0-2
|
|
|
Allele name and predicted product
|
54a
54b |
agggcttttctggg<2747bp deletion>ggagagg null
|
| |
|
突变创建时间 |
2015-04-18
|
|
填表时间 |
2013/12/17
|
|
修改时间 |
2016/12/30
|
|
PI/负责人 (工作单位) |
|
