ZKO number |
ZKO969 |
Recent progress of this line |
F2 individuals have been indentified
|
Gene name |
gstcd
|
Ensembl ID |
ENSDARG00000036494
|
Zebrafish strain |
AB
|
Cas9 target site 1 sequence |
sense strand
GCCAGTTGATGCCCGACCGCAggg
|
Cas9 target site 2 sequence |
|
Position in the gene |
Exon 2
|
Restriction enzyme (RE) |
Bsl I (CCNN/NNNNNGG)
|
PCR primer (forward) |
TGTCCTACAGCGAATGCTC
|
PCR primer (reverse) |
ctaagcatttccagcacctgt
|
Length of expected PCR product (bp)
|
369
|
Length of expected RE digestion product (bp) |
250 bp +119 bp
|
Plasmids used for making Cas9 mRNA |
pXT7-hCAS9
|
Plasmids?used?for?making?gRNA |
pMD19T-gRNA
|
Primers?used?for?making?gRNA |
Pr-ZKO 969
|
Dosage of injected Cas9 mRNA (ng/ul)
|
1000
|
Dosage of injected gRNA (ng/ul)
|
300
|
Survival rate of F0 embryo @ 1dpf (%)
|
52
|
Method to estimate mutation efficiency |
Restriction enzyme digestion68
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO969.pdf
|
Mutation efficiency in F0 embryos (%)
|
30
|
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture) |
|
Mutation type
|
deletion
|
Germline transmission efficiency in adult F0
|
6/20
|
Wild-type (WT) sequence
|
CCCGACCGCAGGGACGTGGCGTCACTTTT
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
CCCGACC AGGGACGTGGCGTCACTTTT(+0bp,-2bp)
|
F0-2
|
CCCG-------CAGGGACGTGGCG (+0bp,-4bp)
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
CCCGACC AGGGACGTGGCGTCACTTTT(+0bp,-2bp)
|
F0-2
|
CCCG-------CAGGGACGTGGCG (+0bp,-4bp)
|
Allele name and predicted product
|
969a 969b |
GCCAGTTGATGCCCGACC-- AGGGAC truncated protein (128aa)
|
CCCG---CAGGGACGTGGCG truncated protein (153aa) |
突变创建时间 |
2015-03-18
|
填表时间 |
2013/12/17
|
修改时间 |
2016/12/29
|
PI/负责人 (工作单位) |
|