| ZKO number |
ZKO38 |
| Recent progress of this line |
F2 individuals have been indentified
|
|
Gene name |
NFKBIZ
|
|
Ensembl ID |
ENSDARG00000095117
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
5’- GGAGCTGATGAGCCGAGATG agg -3’ (sense strand)
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
Exon 5
|
|
Restriction enzyme (RE) |
T7
|
|
PCR primer (forward) |
5’- GGACAGATGGAGTGTGTGTCCAG-3’
|
|
PCR primer (reverse) |
5’- GTGTGTGTACTGACGTGTCTCCG -3’
|
|
Length of expected PCR product (bp)
|
325
|
|
Length of expected RE digestion product (bp) |
301+24
|
|
Plasmids used for making Cas9 mRNA |
pSP6-2sNLS-spCas9
|
| Plasmids?used?for?making?gRNA |
pMD19T-gRNA
|
| Primers?used?for?making?gRNA |
Pr-ZKO 38
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
300
|
|
Dosage of injected gRNA (ng/ul)
|
50
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
85.9
|
|
Method to estimate mutation efficiency |
T7EI detection
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO38.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
75
|
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture) |
|
|
Mutation type
|
deletion
|
|
Germline transmission efficiency in adult F0
|
13.33%
|
|
Wild-type (WT) sequence
|
ATGGCAGATCCAGCAGGAGGATGAGAAACTGTCTGCGTTGAGCCCTGCGGAGCTGATGAGCCGAGATGAGGACGGAGACAC
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
ATGGCAGATCCAGC----AGGACGGAGACAC（-54bp）
|
|
F0-2
|
ATGGCAGATCCA----AGGACGGAGACAC (-21bp,+7 bp)
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
|
|
F0-2
|
ATGGCAGATCCA----AGGACGGAGACAC (-21bp,+7 bp)
|
|
Allele name and predicted product
|
38a
38b |
|
| ATGGCAGATCCA----AGGACGGAGACAC ;Truncated protein (223 aa) |
|
突变创建时间 |
|
|
填表时间 |
2013/12/24
|
|
修改时间 |
2016/8/8
|
|
PI/负责人 (工作单位) |
|
