| ZKO number |
ZKO445 |
| Recent progress of this line |
F2 generation has been raised
|
|
Gene name |
smim14
|
|
Ensembl ID |
ENSDARG00000005799
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GGCCCCAACTCCTCAGTAGAagg
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
Exon 4
|
|
Restriction enzyme (RE) |
DdeI(C︱TNAG)
|
|
PCR primer (forward) |
GTAAGTGTGTCAGCCCTCCAAAATC
|
|
PCR primer (reverse) |
TGCACTGCAGCACTTGCACTGACT
|
|
Length of expected PCR product (bp)
|
372
|
|
Length of expected RE digestion product (bp) |
113+259
|
|
Plasmids used for making Cas9 mRNA |
pGH-T7-zCAS9
|
| Plasmids used for making gRNA |
pMD19T-gRNA
|
| Primers used for making gRNA |
Pr-ZKO
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
500
|
|
Dosage of injected gRNA (ng/ul)
|
200
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
71.4
|
|
Method to estimate mutation efficiency |
Restriction enzyme digestionDdeI(C︱TNAG)
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO445.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
54.8
|
Picture from F0 embryos
(gel picture or sequencing picture) |
|
|
Mutation type
|
Indel
|
|
Germline transmission efficiency in adult F0
|
55.6%
|
|
Wild-type (WT) sequence
|
GGCCCCAACTCCTCAGTAGAAGG
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
|
|
F0-2
|
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
GGCCCCAACTCCTCGACATCACTATACCCATGAAGG（-4bp,+17bp）
|
|
F0-2
|
GGCCCCAACTCCTGAAGGTTGGG（-8bp,+8bp introduced a stop codon TGA）
|
|
Allele name and predicted product
|
445a
445b |
GGCCCCAACTCCTCGACATCACTATACCCATGAAGG Truncated protein(53aa)
|
| GGCCCCAACTCCTGAAGGTTGGG Truncated protein(47aa) |
|
突变创建时间 |
2014-09-02
|
|
填表时间 |
2013/12/17
|
|
修改时间 |
2016/8/12
|
|
PI/负责人 (工作单位) |
|