| ZKO number |
ZKO1066 |
| Recent progress of this line |
F1 individuals have been identified
|
|
Gene name |
cct4
|
|
Ensembl ID |
ENSDARG00000013475
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GAACAAGTCTGGGCCCTAA
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
Exon 2
|
|
Restriction enzyme (RE) |
Apa I (GGGCC|C)
|
|
PCR primer (forward) |
GGATTGGTGGCTAATGATGA
|
|
PCR primer (reverse) |
ACATTCCTCTTGTAGTGCTT
|
|
Length of expected PCR product (bp)
|
471
|
|
Length of expected RE digestion product (bp) |
72+399
|
|
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)
|
300
|
|
Dosage of injected gRNA (ng/ul)
|
50
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
91.6
|
|
Method to estimate mutation efficiency |
Restriction enzyme digestionRE digestion (BstYI)
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO1066.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
20
|
Picture from F0 embryos
(gel picture or sequencing picture) |
|
|
Mutation type
|
Indel
|
|
Germline transmission efficiency in adult F0
|
20% (3/15)
|
|
Wild-type (WT) sequence
|
tgccatcagaacaagtctgggccctaagggcatggataaaatggtatgttt
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
|
|
F0-2
|
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
50% (m/n: Number of F1 carrier / Total number of F1 screened)
tgccatcagaacaagtctgggcc---------tggataaaatggtatgtt (+0 bp, -9bp)
|
|
F0-2
|
27.3% (m/n: Number of F1 carrier / Total number of F1 screened)
tgccatcagaacaa-------------gggcatggataaaatggtatgtt (+0 bp, -13 bp)
|
|
Allele name and predicted product
|
1066a
1066b |
rdkpaqirysninagkavadairtslg---ldkmiqdgkgdvtitndgatilk; Truncated protein (530aa)
|
| rdkpaqirysninagkavadairtrawik*; Truncated protein (79 aa) |
|
突变创建时间 |
2014-02-15
|
|
填表时间 |
2014/1/4
|
|
修改时间 |
2016/11/11
|
|
PI/负责人 (工作单位) |
|