ZKO number |
ZKO1020 |
Recent progress of this line |
F1 progeny has been raised
|
Gene name |
asna1
|
Ensembl ID |
ENSDARG00000018190
|
Zebrafish strain |
AB
|
Cas9 target site 1 sequence |
sense strand
GGAGGCCATGAGTGCTTTCCCtgg
|
Cas9 target site 2 sequence |
|
Position in the gene |
exon3
|
Restriction enzyme (RE) |
CC/TGG(Bst NI)
|
PCR primer (forward) |
TGTCTGTCTGCAGGAGATCG
|
PCR primer (reverse) |
CTTGAACGCGAGTCTGACCT
|
Length of expected PCR product (bp)
|
180
|
Length of expected RE digestion product (bp) |
123+57
|
Plasmids used for making Cas9 mRNA |
pXT7-hCAS9
|
Plasmids?used?for?making?gRNA |
pMD19T-gRNA
|
Primers?used?for?making?gRNA |
Pr-ZKO 1020
|
Dosage of injected Cas9 mRNA (ng/ul)
|
300
|
Dosage of injected gRNA (ng/ul)
|
50
|
Survival rate of F0 embryo @ 1dpf (%)
|
80
|
Method to estimate mutation efficiency |
Restriction enzyme digestionenzyme
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO1020.pdf
|
Mutation efficiency in F0 embryos (%)
|
94
|
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture) |
|
Mutation type
|
Indel
|
Germline transmission efficiency in adult F0
|
66.6
|
Wild-type (WT) sequence
|
GGAGGCCATGAGTGCTTTCCCTGGCATCGATGAGGCCATGAGCTACGCAGAGGTCA
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
ATGGGGAAAAAGATGATGCAGGAGGCCATGGCCATGAGCTACGCAGAGGTCATGAGgtca(-13bp)
|
F0-2
|
GGCCATGAGTGCTTTGGCATCGATGAGGCC(-4bp)
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
ATGGGGAAAAAGATGATGCAGGAGGCCATGGCATCGATGAGGCCATGAGCTACGCAGAGG(-13bp)
|
F0-2
|
GGCCATGAGTGCTTTGGCATCGATGAGGCC(-4bp)
|
Allele name and predicted product
|
1020a 1020b |
ATGGGGAAAAAGATGATGCAGGAGGCCATGGCATCGATGAGGCCATGAGCTACGCAGAGG(-13bp);truncated protein,135aa
|
GGCCATGAGTGCTTTGGCATCGATGAGGCC(-4bp);truncated protein,138aa |
突变创建时间 |
2014-09-03
|
填表时间 |
2013/12/23
|
修改时间 |
2016/10/24
|
PI/负责人 (工作单位) |
|