ZKO number |
ZKO279 |
Recent progress of this line |
F2 individuals have been indentified
|
Gene name |
CR388002.2
|
Ensembl ID |
ENSDARG00000088299
|
Zebrafish strain |
AB
|
Cas9 target site 1 sequence |
antisense strand
GGATTTTCACAGCACTCATTagg
|
Cas9 target site 2 sequence |
|
Position in the gene |
Exon 5
|
Restriction enzyme (RE) |
n/a
|
PCR primer (forward) |
TTCCGCCTAATCCTATACATCAT
|
PCR primer (reverse) |
CTCCTACAGGTTCTGGGTCTTCT
|
Length of expected PCR product (bp)
|
554
|
Length of expected RE digestion product (bp) |
|
Plasmids used for making Cas9 mRNA |
pXT7-hCAS9
|
Plasmids?used?for?making?gRNA |
pT7-gRNA-ZKO279
|
Primers?used?for?making?gRNA |
Pr-ZKO 279
|
Dosage of injected Cas9 mRNA (ng/ul)
|
200
|
Dosage of injected gRNA (ng/ul)
|
25
|
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/ZKO279.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
|
3/8
|
Wild-type (WT) sequence
|
GGATTTTCACAGCACTCATTaggTCAGACA
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
|
F0-2
|
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
GGATT-------------TTAGGTCAGACA(-13bp)
|
F0-2
|
GGATTTTCACAGCAC--------TCAGACA(-8bp)
|
Allele name and predicted product
|
279a 279b |
GGATT-------------TTAGGTCAGACA Truncated protein(204aa)
|
GGATTTTCACAGCAC--------TCAGACA Truncated protein(201aa) |
突变创建时间 |
2014-10-23
|
填表时间 |
2013/12/28
|
修改时间 |
2016/8/11
|
PI/负责人 (工作单位) |
|