| ZKO number |
ZKO762 |
| Recent progress of this line |
F1 individuals have been identified
|
|
Gene name |
calmegin
|
|
Ensembl ID |
ENSDARG00000009315
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
ATGGGAGGTTGAGCCCCTGAagg
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
exon 5
|
|
Restriction enzyme (RE) |
n/a
|
|
PCR primer (forward) |
TGTGTGGAACCAAGCATTTT
|
|
PCR primer (reverse) |
CGCCACAGTCAATTCCATC
|
|
Length of expected PCR product (bp)
|
383
|
|
Length of expected RE digestion product (bp) |
|
|
Plasmids used for making Cas9 mRNA |
pGH-T7-zCAS9
|
| Plasmids?used?for?making?gRNA |
pMD19T-gRNA
|
| Primers?used?for?making?gRNA |
Pr-ZKO 762
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
600
|
|
Dosage of injected gRNA (ng/ul)
|
1000
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
85
|
|
Method to estimate mutation efficiency |
sequencing of PCR product
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO762.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
10
|
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture) |
|
|
Mutation type
|
Indel
|
|
Germline transmission efficiency in adult F0
|
50
|
|
Wild-type (WT) sequence
|
tgtgtggaaccaagcatttttacagtgtatcataggtcatggaaattcagctttttggttcgtgaaagtctggtaatttgagtgggaaccttttcattgtctgtagGTAAATGGGAGGTTGAGCCCCTGAAGGAAAATACAGTGCCTGGTGATTTAGGTTTGGTGCTGAAATCTCGTGCCAAGCATCATGCAATCTCGGCTCTGCTTAACAAACCATTTGTGTTTAAGGACAAACCTCTCATCGTCCAgtaagtgtgttttgtatttgttgtaatctgacatttgacataccttggatttctttaataaaacttgttaattctaacaaaacccaatctatatacagGTACGAGGTCAATTTTCAAGATGGAATTGACTGTGGCG
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
|
|
F0-2
|
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
>WT
ATTTAGGTTTGGTGCTGAAATCTCGTGCCAAGCATCATGCAATCTCGGCTCTGCTTAACAAACCATTTGTGTTTAAGGACAAACCTCTCATCGTCCAgtaagtgt
>ZKO762b
ATTTAGGTTTGGTGCTGAAATCTCGTGCC----------------------------------------------AGGACAAACCTCTCATCGTCCAgtaagtgt
|
|
F0-2
|
>WT
accttttcattgtctgtagGTAAATGGGAG-----------------------------GTTGAGCCCCTGAAGGAAAATACAGTGCCT
>ZKO762b
accttttcattgtctgtagGTAAATGGGAGATTTCTTTATGATTGATATTTTGTCTGCATAAAAAGTAAATATTTTTGTTGAGCCCCTGAAGGAAAATACAGTGCCT
|
|
Allele name and predicted product
|
762a
762b |
ATTTAGGTTTGGTGCTGAAATCTCGTGCC----------------------------------------------AGGACAAACCTCTCATCGTCCA truncated protein (159aa)
|
| accttttcattgtctgtagGTAAATGGGAGATTTCTTTATGATTGATATTTTGTCTGCATAAAAAGTAAATATTTTTGTTGAGCCCCTGAAGGAAAATACAGTGCCT truncated protein (109aa) |
|
突变创建时间 |
2013-12-16
|
|
填表时间 |
2013/12/18
|
|
修改时间 |
2017/1/5
|
|
PI/负责人 (工作单位) |
|
