| ZKO number |
ZKO111 |
| Recent progress of this line |
F2 individuals have been indentified
|
|
Gene name |
hecw2b
|
|
Ensembl ID |
ENSDARG00000063253
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GGGGCAGAGGGGCAGGTGGAtgg
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
exon4
|
|
Restriction enzyme (RE) |
BtsCI
|
|
PCR primer (forward) |
agcatttcgttcaagtcttttgt
|
|
PCR primer (reverse) |
TATATGCGGCGGTTCATTCC
|
|
Length of expected PCR product (bp)
|
443
|
|
Length of expected RE digestion product (bp) |
350+90
|
|
Plasmids used for making Cas9 mRNA |
pGH-T7-zCAS9
|
| Plasmids used for making gRNA |
pMD19T-gRNA
|
| Primers used for making gRNA |
Pr-ZKO 111
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
500
|
|
Dosage of injected gRNA (ng/ul)
|
100
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
80
|
|
Method to estimate mutation efficiency |
Restriction enzyme digestionBtsCI digestion
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO111.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
40
|
Picture from F0 embryos
(gel picture or sequencing picture) |
|
|
Mutation type
|
Indel
|
|
Germline transmission efficiency in adult F0
|
30%
|
|
Wild-type (WT) sequence
|
AATAGTGTGTCATGGTGTTCTCTGCAGGTTGGGGCAGAGGGGCAGGTGGATGGACAGTCTGTCACTGAACACTGCAGGA A
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
AATAGTGTGTCATGGTGTTCTCTGCAGGTTGGGGCAGAGGGG--------------------CACTGAACACTGCAGGAA -20bp
|
|
F0-2
|
AATAGTGTGTCATGGTGTTCTCTGCAGGTTGGGGCAGAGGGGCAGG-------ACAGTCTGGGACTGAACACTGCAGGAA
-7bp
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
AATAGTGTGTCATGGTGTTCTCTGCAGGTTGGGGCAGAGGGGCAGG-------ACAGTCTGGGACTGAACACTGCAGGAA
-7bp
|
|
F0-2
|
AATAGTGTGTCATGGTGTTCTCTGCAGGTTGGGGCAGAGGGGCAGGTGGA-ctGACAGTCTGTCACTGAACACTGCAGGAA -1bp
|
|
Allele name and predicted product
|
111a
111b |
GGGGCAGAGGGGCAGG------- truncated protein(196aa)
|
| |
|
突变创建时间 |
|
|
填表时间 |
2013/12/17
|
|
修改时间 |
2016/12/30
|
|
PI/负责人 (工作单位) |
|