| ZKO number |
ZKO108 |
| Recent progress of this line |
F2 generation has been raised
|
|
Gene name |
zgc:153922
|
|
Ensembl ID |
ENSDARG00000018008
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GGAGGACACCTGCGCATCTATtgg
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
Exon 3
|
|
Restriction enzyme (RE) |
Fsp I (TGC/GCA)
|
|
PCR primer (forward) |
5’- GCCCGATGACTGGAAGgt -3’…………51.9
|
|
PCR primer (reverse) |
5’- agcgacagatgtgccaatag -3’…………50.2
|
|
Length of expected PCR product (bp)
|
510
|
|
Length of expected RE digestion product (bp) |
264 bp +246 bp
|
|
Plasmids used for making Cas9 mRNA |
pXT7-hCAS9
|
| Plasmids used for making gRNA |
pMD19T-gRNA
|
| Primers used for making gRNA |
Pr-ZKO 108
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
1000
|
|
Dosage of injected gRNA (ng/ul)
|
300
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
60
|
|
Method to estimate mutation efficiency |
Restriction enzyme digestion20
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO108.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
|
20
|
|
Wild-type (WT) sequence
|
AAATGGCTGGAGGACACCTGCGCATCTATTGGCTGGAACACTCAA
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
ACCTGCGC TGGAAGGTGGC TGGCTGGAAACA
（+11bp,-5bp）
|
|
F0-2
|
AAA------A-------------------------------GGCTGGAACACT
（-27bp，+1bp）
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
ACCTGCGC TGGAAGGTGGC TGGCTGGAAACA
（+11bp,-5bp）
|
|
F0-2
|
AAA------A-------------------------------GGCTGGAACACT
（-27bp，+1bp）
|
|
Allele name and predicted product
|
108a
108b |
ACCTGCGC TGGAAGGTGGC TGGCTGGAAACA truncated protein（78aa）
|
| AAA------A-------------------------------GGCTGGAACACT truncated protein (68aa) |
|
突变创建时间 |
|
|
填表时间 |
2013/12/17
|
|
修改时间 |
2016/12/29
|
|
PI/负责人 (工作单位) |
|