| ZKO number |
ZKO59 |
| Recent progress of this line |
F2 individuals have been indentified
|
|
Gene name |
atp1a1a.2
|
|
Ensembl ID |
ENSDARG00000007739
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GGGACTTGTGCTTGCGTTTGtgg
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
exon5
|
|
Restriction enzyme (RE) |
n/a
|
|
PCR primer (forward) |
TTCAAACGGAAAGTGCAACCC
|
|
PCR primer (reverse) |
GCAGGGATTCTGTCTCCACC
|
|
Length of expected PCR product (bp)
|
408
|
|
Length of expected RE digestion product (bp) |
|
|
Plasmids used for making Cas9 mRNA |
pXT7-hCAS9
|
| Plasmids used for making gRNA |
pMD19T-gRNA
|
| Primers used for making gRNA |
Pr-ZKO 59
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
300
|
|
Dosage of injected gRNA (ng/ul)
|
100
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
75
|
|
Method to estimate mutation efficiency |
sequencing of PCR product
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO59.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
70
|
Picture from F0 embryos
(gel picture or sequencing picture) |
|
|
Mutation type
|
deletion
|
|
Germline transmission efficiency in adult F0
|
30%(3/10)
|
|
Wild-type (WT) sequence
|
CTGTACCTGGGACTTGTGCTTGCGTTTGTGGTCATAGT
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
|
|
F0-2
|
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
31.25%(5/16)
CTGTACCTGGGACTTGTG------------GTCATAGT(-12)
|
|
F0-2
|
31.25%(5/16)
CTGTACCTGGGACTTGTGCTTGCGACTTGTGGTCATGACTTGTGGTCATAGT(-1,+15)
|
|
Allele name and predicted product
|
59a
59b |
CTGTACCTGGGACTTGTG------------GTCATAGT(-12),delete aa136-139
|
| CTGTACCTGGGACTTGTGCTTGCGACTTGTGGTCATGACTTGTGGTCATAGT(-1,+15),truncated protein(145aa) |
|
突变创建时间 |
|
|
填表时间 |
2013/12/18
|
|
修改时间 |
2016/8/11
|
|
PI/负责人 (工作单位) |
|