| ZKO number |
ZKO64 |
| Recent progress of this line |
F1 individuals have been identified
|
|
Gene name |
si:ch211-132g1.6
|
|
Ensembl ID |
ENSDARG00000093102
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GCCAATGGCCAACAGATTGggg
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
exon 2
|
|
Restriction enzyme (RE) |
BstX 1
|
|
PCR primer (forward) |
AAAAAGCTGCAGATATCAGAT
|
|
PCR primer (reverse) |
GTGCATTCACGCTTCACAGT
|
|
Length of expected PCR product (bp)
|
393
|
|
Length of expected RE digestion product (bp) |
201+192
|
|
Plasmids used for making Cas9 mRNA |
pXT7-hCAS9
|
| Plasmids used for making gRNA |
pMD19T-gRNA
|
| Primers used for making gRNA |
Pr-ZKO 64
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
1000
|
|
Dosage of injected gRNA (ng/ul)
|
300
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
70
|
|
Method to estimate mutation efficiency |
sequencing of PCR product
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO64.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
50
|
Picture from F0 embryos
(gel picture or sequencing picture) |
|
|
Mutation type
|
Indel
|
|
Germline transmission efficiency in adult F0
|
20%
|
|
Wild-type (WT) sequence
|
ACCGGCACATATAAATACAAAGCTTTTAATGCCAATGGCCAACAGATTGGGGATGAAACTGTGGAACTGAAAGTTTATGGTG
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
ACCGGCACATATAAATACAAAGCTTTTAATGCCAATGGCCAACAGAGATGAAAC-TGGGGATGAAACTGTGGAACTGAAAGTTTATGGTG
(-1bp,+8bp)
|
|
F0-2
|
ACCGGCACATATAAATACAAAGCTTTTAATGCCAATGGCC-----ATTGGGGATGAAACTGTGGAACTGAAAGTTTATGGTG
(-5bp)
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
30%
ACCGGCACATATAAATACAAAGCTTTTAATGCCAATGGCCAACAGAGATGAAAC-TGGGGATGAAACTGTGGAACTGAAAGTTTATGGTG
(-1bp,+8bp)
|
|
F0-2
|
30%
ACCGGCACATATAAATACAAAGCTTTTAATGCCAATGGCC-----ATTGGGGATGAAACTGTGGAACTGAAAGTTTATGGTG
(-5bp)
|
|
Allele name and predicted product
|
64a
64b |
AATGCCAATGGCCAACAGAGATGAAAC-TGGGGATGAAACTGTG Truncated protein(109aa)
|
| AATGCCAATGGCC-----ATTGGGGATGAAACTGTGGAACTGAAAG Truncated protein(109aa) |
|
突变创建时间 |
2015-06-08
|
|
填表时间 |
2013/12/16
|
|
修改时间 |
2016/12/29
|
|
PI/负责人 (工作单位) |
|