| ZKO number |
ZKO448 |
| Recent progress of this line |
F2 generation has been raised
|
|
Gene name |
n4bp2
|
|
Ensembl ID |
ENSDARG00000061451
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GGATCCAGAAGTCATTTGCAtgg
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
Exon 1
|
|
Restriction enzyme (RE) |
MslI(CAYNN︱NNRTG)
|
|
PCR primer (forward) |
GACATAGTCCGTCCCGAGCACC
|
|
PCR primer (reverse) |
CAATTGTATGCAGCCTTGAGACG
|
|
Length of expected PCR product (bp)
|
373
|
|
Length of expected RE digestion product (bp) |
219+154
|
|
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 (%)
|
80
|
|
Method to estimate mutation efficiency |
Restriction enzyme digestionMslI(CAYNN︱NNRTG)
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO448.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
37.6
|
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture) |
|
|
Mutation type
|
Indel
|
|
Germline transmission efficiency in adult F0
|
40%
|
|
Wild-type (WT) sequence
|
GGATCCAGAAGTCATTTGCATGG
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
|
|
F0-2
|
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
GGATCCAGAAGTCATTTGGTGCTCTCAGAGGAAGCATGG（+16bp）
|
|
F0-2
|
GGATCCAGAAGTCATT-GCATGG（-1bp）
|
|
Allele name and predicted product
|
448a
448b |
GGATCCAGAAGTCATTTGGTGCTCTCAGAGGAAGCATGG Truncated protein(94aa)
|
| GGATCCAGAAGTCATT-GCATGGTruncated protein(107aa) |
|
突变创建时间 |
2014-09-02
|
|
填表时间 |
2013/12/17
|
|
修改时间 |
2016/8/12
|
|
PI/负责人 (工作单位) |
|
