| ZKO number |
ZKO422 |
| Recent progress of this line |
F2 generation has been raised
|
|
Gene name |
dnah7l
|
|
Ensembl ID |
ENSDARG00000000606
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GGACATCCCAGGAGCCAGAGagg
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
exon 2
|
|
Restriction enzyme (RE) |
NIaIV(GGN | NCC)
|
|
PCR primer (forward) |
GTATCGCTGATTGGCTGTAG
|
|
PCR primer (reverse) |
CAGAGAAGGAGGGACAGTTG
|
|
Length of expected PCR product (bp)
|
473
|
|
Length of expected RE digestion product (bp) |
381+92
|
|
Plasmids used for making Cas9 mRNA |
pXT7-hCAS9
|
| 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 (%)
|
|
|
Method to estimate mutation efficiency |
Restriction enzyme digestionNIaIV (GGN | NCC)
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO422.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
53
|
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture) |
|
|
Mutation type
|
deletion
|
|
Germline transmission efficiency in adult F0
|
78%(7/9: Number of positive F0 / Total number of F0 screened)
|
|
Wild-type (WT) sequence
|
AGAAAGGACATCCCAGGAGCCAGAGAGGGCCAG
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
|
|
F0-2
|
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
AGAAAGGACATCCCAGGAG-AGGAGAGGGCCAG-3bp,+2bp)
|
|
F0-2
|
AGAAAGGACATCCCAGGAG--AAGGAGGGCCAG(-5bp,+3bp)
|
|
Allele name and predicted product
|
422a
422b |
AGAAAGGACATCCCAGGAG-AGGAGAGGGCCAG ; Truncated protein(48aa)
|
| AGAAAGGACATCCCAGGAG--AAGGAGGGCCAG; Truncated protein(27aa) |
|
突变创建时间 |
2014-09-01
|
|
填表时间 |
2013/12/17
|
|
修改时间 |
2016/8/12
|
|
PI/负责人 (工作单位) |
|
