| ZKO number |
ZKO275 |
| Recent progress of this line |
F2 individuals have been indentified
|
|
Gene name |
pcdh18a
|
|
Ensembl ID |
ENSDARG00000089805
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GGACTGAGACAGCACAACAAagg
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
exon 1
|
|
Restriction enzyme (RE) |
n/a
|
|
PCR primer (forward) |
CCAGACGGAGGTTGAGCTTACAG
|
|
PCR primer (reverse) |
CACGAGCAGAGCAAGTTTGAGTA
|
|
Length of expected PCR product (bp)
|
353
|
|
Length of expected RE digestion product (bp) |
|
|
Plasmids used for making Cas9 mRNA |
pXT7-hCAS9
|
| Plasmids?used?for?making?gRNA |
pT7-gRNA-ZKOpT7-gRNA-ZKO275
|
| Primers?used?for?making?gRNA |
Pr-ZKO 275
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
200
|
|
Dosage of injected gRNA (ng/ul)
|
25
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
80
|
|
Method to estimate mutation efficiency |
sequencing of PCR product
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO275.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
66.7
|
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture) |
|
|
Mutation type
|
deletion
|
|
Germline transmission efficiency in adult F0
|
6/8
|
|
Wild-type (WT) sequence
|
GGACTGAGACAGCACAACAAaggAAGCGAC
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
|
|
F0-2
|
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
GGAAAGGAAGCGACAGG----ggAAGCGAC(-18bp,+14bp)
|
|
F0-2
|
GGACTGAGACAG-----CAAaggAAGCGAC(-5bp)
|
|
Allele name and predicted product
|
275a
275b |
GGAAAGGAAGCGACAGG----ggAAGCGAC Delete 18bp, insert 14bp in 5'-UTR
|
| GGACTGAGACAG-----CAAaggAAGCGAC Delete 5bp in 5'-UTR |
|
突变创建时间 |
2014-08-20
|
|
填表时间 |
2013/12/28
|
|
修改时间 |
2016/8/11
|
|
PI/负责人 (工作单位) |
|
