| ZKO number |
ZKO835 |
| Recent progress of this line |
F2 individuals have been indentified
|
|
Gene name |
cryba1l2
|
|
Ensembl ID |
ENSDARG00000014803
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
antisense strand
GGAAATGCTCCTGCTCGAACA
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
Exon3
|
|
Restriction enzyme (RE) |
TaqI
|
|
PCR primer (forward) |
AGCAGAGGATTGTGTAGGAG
|
|
PCR primer (reverse) |
TTTGCATTATAGCAGCTCGGT
|
|
Length of expected PCR product (bp)
|
398
|
|
Length of expected RE digestion product (bp) |
126bp +272bp
|
|
Plasmids used for making Cas9 mRNA |
pGH-T7-zCAS9
|
| Plasmids?used?for?making?gRNA |
pMD19T-gRNA
|
| Primers?used?for?making?gRNA |
Pr-ZKO 835
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
200
|
|
Dosage of injected gRNA (ng/ul)
|
150
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
90
|
|
Method to estimate mutation efficiency |
Restriction enzyme digestionTaqI
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO835.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
|
3.33%
|
|
Wild-type (WT) sequence
|
TGAAACCGTGGCATGTGCACGTGCACGTGTGTGTGTGTGTGTGTGTGTGTGTTTTCAGGTGACCGTGTTCGAGCAGGAGCATTTCCAAGGCAAGTGTCA
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
TGAAACCGTGGCATGTGCACGTGCACGTGTGTGTGTGTGTGTGTGTGTGTGTTTTCAGGTGACCGTGTCAGGTGACCGTACGAGCAGGAGCATTTCCAAGGCAAGTGTCA （+12，-1）
|
|
F0-2
|
TGAAACCGTGGCATGTGCACGTGCACA------TGTGTGTGTGTGTGTGTGTTTTCAGGTGACCGTGTCAGGTGACCGTCCGAGCAGGAGCATTTCCAAGGCAAGTGTCA（+15，-10）
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
TGAAACCGTGGCATGTGCACGTGCACGTGTGTGTGTGTGTGTGTGTGTGTGTTTTCAGGTGACCGTGTCAGGTGACCGTACGAGCAGGAGCATTTCCAAGGCAAGTGTCA （+12，-1）
|
|
F0-2
|
TGAAACCGTGGCATGTGCACGTGCACA------TGTGTGTGTGTGTGTGTGTTTTCAGGTGACCGTGTCAGGTGACCGTCCGAGCAGGAGCATTTCCAAGGCAAGTGTCA（+15，-10）
|
|
Allele name and predicted product
|
835a
835b |
（+12，-1）
|
| （+15，-10） |
|
突变创建时间 |
2014-08-23
|
|
填表时间 |
2013/12/19
|
|
修改时间 |
2017/3/21
|
|
PI/负责人 (工作单位) |
|
