| ZKO number |
ZKO1058 |
| Recent progress of this line |
F1 individuals have been identified
|
|
Gene name |
zgc:66455
|
|
Ensembl ID |
ENSDARG00000052375
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GATCCACATGCTTGCCCGG
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
Exon 3
|
|
Restriction enzyme (RE) |
Sma1/Xma1 (C︱CCGGG)
|
|
PCR primer (forward) |
gtgaatgttgcattaatgttgtg
|
|
PCR primer (reverse) |
gctctgtagcgtgcatagaag
|
|
Length of expected PCR product (bp)
|
571
|
|
Length of expected RE digestion product (bp) |
129+403
|
|
Plasmids used for making Cas9 mRNA |
pXT7-hCAS9
|
| Plasmids?used?for?making?gRNA |
pMD19T-gRNA
|
| Primers?used?for?making?gRNA |
Pr-ZKO 1058
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
300
|
|
Dosage of injected gRNA (ng/ul)
|
50
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
91.6
|
|
Method to estimate mutation efficiency |
sequencing of PCR product
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO1058.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
|
50% (2/4)
|
|
Wild-type (WT) sequence
|
CCCCCGGGCAAGCATGTGGGATC
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
CCCCCGGGGCAAGCATGTGGATC ( +1 bp)
CCCCCG--CAAGCATGTGGATC ( -2 bp)
CCCCCG---AAGCATGTGGATC (-3 bp)
|
|
F0-2
|
CCCCC--CTGCAACTGCAAGCATGTGGATC (+8 bp, -2 bp)
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
25% (5/20)
CCCCCGGGGCAAGCATGTGGATC (+1 bp)
|
|
F0-2
|
F1-2 10% (2/20)
CCCCCG--CAAGCATGTGGATC ( -2 bp)
|
|
Allele name and predicted product
|
1058a
1058b |
CCCCCGGGGCAAGCATGTGGATC; (+1) ;Truncated protein (111aa)
|
| CCCCCG--CAAGCATGTGGATC ( -2 bp);Truncated protein (110aa) |
|
突变创建时间 |
2013-09-10
|
|
填表时间 |
2014/1/4
|
|
修改时间 |
2016/11/11
|
|
PI/负责人 (工作单位) |
|
