| ZKO number |
ZKO105 |
| Recent progress of this line |
F2 individuals have been indentified
|
|
Gene name |
cnppd1
|
|
Ensembl ID |
ENSDARG00000027078
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GCTCTATCATGCCCCATGACagg
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
Exon 2
|
|
Restriction enzyme (RE) |
Bsl I (CCNN/NNNNNGG)
|
|
PCR primer (forward) |
5’- tgttactgaacgaatgtgcga -3’…………51.7
|
|
PCR primer (reverse) |
5’- ggacaacagtatctcccgcat -3’…………53.0
|
|
Length of expected PCR product (bp)
|
608
|
|
Length of expected RE digestion product (bp) |
266 bp + 342 bp
|
|
Plasmids used for making Cas9 mRNA |
pXT7-hCAS9
|
| Plasmids?used?for?making?gRNA |
pMD19T-gRNA
|
| Primers?used?for?making?gRNA |
Pr-ZKO 105
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
1000
|
|
Dosage of injected gRNA (ng/ul)
|
300
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
75
|
|
Method to estimate mutation efficiency |
Restriction enzyme digestionBsl 1
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO105.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
30
|
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture) |
|
|
Mutation type
|
Indel
|
|
Germline transmission efficiency in adult F0
|
30% 3/10
|
|
Wild-type (WT) sequence
|
ACAGCACGCTGGATGCTCTATCATGCCCCATGACAGgtgagtcaacttaatttaaaaataggcatgg
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
ACGCTGGATGCTCTATCATGCCGTGACAGgtgagtcaacttaatttaaaaataggcatgg (+1bp, -3 bp)
|
|
F0-2
|
ACAGCACGCTGGATGCTCTATCATGCCTAAGTCGACAGgtgagtcaacttaatttaaaaataggcatgg (-4bp, +6bp)
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
ACGCTGGATGCTCTATCATGCCGTGACAGgtgagtcaacttaatttaaaaataggcatgg (+1bp, -3 bp)
|
|
F0-2
|
ACAGCACGCTGGATGCTCTATCATGCCTAAGTCGACAGgtgagtcaacttaatttaaaaataggcatgg (-4bp, +6bp)
|
|
Allele name and predicted product
|
105a
105b |
ATCATGCC--GTGACAG truncated protein（69aa）
|
| ATCATGCCTAAGTCGACAG（59aa） |
|
突变创建时间 |
|
|
填表时间 |
2013/12/16
|
|
修改时间 |
2016/12/29
|
|
PI/负责人 (工作单位) |
|
