ZKO number |
ZKO100 |
Recent progress of this line |
F2 individuals have been indentified
|
Gene name |
nrp2a
|
Ensembl ID |
ENSDARG00000031622
|
Zebrafish strain |
AB
|
Cas9 target site 1 sequence |
antisense strand
GCCACAGTATCGTCCGATCAAggg
|
Cas9 target site 2 sequence |
|
Position in the gene |
Exon2
|
Restriction enzyme (RE) |
Bsl I (CCNNNNN/NNGG)
|
PCR primer (forward) |
5’- ctcgggatcataggagcagt -3’…………50.7
|
PCR primer (reverse) |
5’- tcatgggaccttcaaatgtg -3’…………50.4
|
Length of expected PCR product (bp)
|
437
|
Length of expected RE digestion product (bp) |
214bp +223 bp
|
Plasmids used for making Cas9 mRNA |
pXT7-hCAS9
|
Plasmids?used?for?making?gRNA |
pMD19T-gRNA
|
Primers?used?for?making?gRNA |
Pr-ZKO 100
|
Dosage of injected Cas9 mRNA (ng/ul)
|
1000
|
Dosage of injected gRNA (ng/ul)
|
300
|
Survival rate of F0 embryo @ 1dpf (%)
|
75.6
|
Method to estimate mutation efficiency |
Restriction enzyme digestion60
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO100.pdf
|
Mutation efficiency in F0 embryos (%)
|
50
|
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture) |
|
Mutation type
|
Indel
|
Germline transmission efficiency in adult F0
|
50% (5/10)
|
Wild-type (WT) sequence
|
tccaacatttccttgtgttcagtGGGACCCTTGATCGGACGATACTGTGGCTCGAAGATCCCTCCAAAGATCCAGTCCTCC
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
tccaacatttccttgtgttcagtGGGACCCTTTGGAGGGACCCTTGAAGATCCCTCCAAAGATCCAGTCCTCC
(+13bp, -21 bp)
sense
|
F0-2
|
tccaacatttccttgtgttcagtGGGACCCTT--------------------------------------------GATCCCTCCAAAGATCCAGTCCTCC
(+0bp, -24bp)
sense
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
tccaacatttccttgtgttcagtGGGACCCTTTGGAGGGACCCTTGAAGATCCCTCCAAAGATCCAGTCCTCC
(+13bp, -21 bp)
sense
|
F0-2
|
tccaacatttccttgtgttcagtGGGACCCTT--------------------------------------------GATCCCTCCAAAGATCCAGTCCTCC
(+0bp, -24bp)
sense
|
Allele name and predicted product
|
100a 100b |
tccaacatttccttgtgttcagtGGGACCCTTTGGAGGGACCCTTGAAGATCCCTCCAAAGATCCAGTCCTCC truncnted protein (108aa)
|
cttgtgttcagtGGGACCCTT--------------------------------------------GATCCCTCCAAAGATCCA functional domain lose protein |
突变创建时间 |
|
填表时间 |
2013/12/16
|
修改时间 |
2016/12/29
|
PI/负责人 (工作单位) |
|