| ZKO number |
ZKO32 |
| Recent progress of this line |
F2 individuals have been indentified
|
|
Gene name |
appa
|
|
Ensembl ID |
ENSDARG00000104279
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GGACCGATATCTGGAGACGCcgg
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
Exon 8
|
|
Restriction enzyme (RE) |
EcoRV
|
|
PCR primer (forward) |
actccctaactgcatcagaacagca
|
|
PCR primer (reverse) |
GCCTGTAGAAAGTGAGCGTGTT
|
|
Length of expected PCR product (bp)
|
494
|
|
Length of expected RE digestion product (bp) |
37+458
|
|
Plasmids used for making Cas9 mRNA |
pSP6-2sNLS-spCas9
|
| Plasmids?used?for?making?gRNA |
pT7-gRNA-ZKOpT7-gRNA-ZKO32
|
| Primers?used?for?making?gRNA |
Pr-ZKO 32
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
300
|
|
Dosage of injected gRNA (ng/ul)
|
80
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
89.7
|
|
Method to estimate mutation efficiency |
T7EI detection
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO32.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
10
|
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture) |
|
|
Mutation type
|
Indel
|
|
Germline transmission efficiency in adult F0
|
50% (m/n: Number of positive F0 / Total number of F0 screened)
|
|
Wild-type (WT) sequence
|
TCCGGATCAGTGGACCGATATCTGGAGACGCCGGCGG
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
TCCxxAxGxAxTxxxxxGxxxxCxxxxxAxxxxxGCGG(-24 bp)
|
|
F0-2
|
TCCGGATGCAGTGGACCGATATCcgatatctCGCCGG(-6bp,+8bp)
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
12.5% (m/n: Number of F1 carrier / Total number of F1 screened)
TCCxxAxGxAxTxxxxxGxxxxCxxxxxAxxxxxGCGG(-24 bp)
|
|
F0-2
|
10% (m/n: Number of F1 carrier / Total number of F1 screened)
TCCGGATGCAGTGGACCGATATCcgatatctCGCCGG(-6bp,+8bp)
|
|
Allele name and predicted product
|
32a
32b |
TCCxxAxGxAxTxxxxxGxxxxCxxxxxAxxxxxGCGG, delete aa358-365
|
| TCCGGATGCAGTGGACCGATATCcgatatctCGCCGG,Truncated protein（392aa） |
|
突变创建时间 |
2014-09-01
|
|
填表时间 |
2013/12/24
|
|
修改时间 |
2016/8/8
|
|
PI/负责人 (工作单位) |
|
