| ZKO number |
ZKO295 |
| Recent progress of this line |
F1 individuals have been identified
|
|
Gene name |
PGBD4
|
|
Ensembl ID |
ENSDARG00000039063
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GGGTCTGTGCCCACTTTATTtgg
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
exon1
|
|
Restriction enzyme (RE) |
n/a
|
|
PCR primer (forward) |
GCGCTGTGAACGCAAGTTAG
|
|
PCR primer (reverse) |
TACAGGTTGCGATTCGGAGG
|
|
Length of expected PCR product (bp)
|
256
|
|
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 295
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
600
|
|
Dosage of injected gRNA (ng/ul)
|
50
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
92
|
|
Method to estimate mutation efficiency |
sequencing of PCR product
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO295.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
15
|
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture) |
|
|
Mutation type
|
Indel
|
|
Germline transmission efficiency in adult F0
|
20%
|
|
Wild-type (WT) sequence
|
AAAAGTCAAAGAAGGGTCTGTGCCCACTTTATTTGGACCAGCTTGCTCCTCCGAATCGCAACCT
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
30%(3/10)
AAAAGTCAAAGAAGGGTCTGTGCCCACTTTxxxxGGACCAGCTTGCTCCTCCGAATCGCAACCT
(-4bp)
|
|
F0-2
|
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
30%(4/12)
AAAAGTCAAAGAAGGGTCTGTGCCCACTTTxxxxGGACCAGCTTGCTCCTCCGAATCGCAACCT
(-4bp)
|
|
F0-2
|
|
|
Allele name and predicted product
|
295a
295b |
AAAAGTCAAAGAAGGGTCTGTGCCCACTTTxxxxGGACCAGCTTGCTCCTCCGAATCGCAACCT;Truncated protein(123aa)
|
| |
|
突变创建时间 |
2013-11-05
|
|
填表时间 |
2013/12/18
|
|
修改时间 |
2016/12/30
|
|
PI/负责人 (工作单位) |
|
