If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO72
Recent progress of this line F1 progeny has been raised
Gene name mbnl2
Ensembl ID ENSDARG00000018460
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GCGTCTTCAGATGTGCCGGtgg
Cas9 target site 2 sequence
Position in the gene exon 2
Restriction enzyme (RE) MSP I
PCR primer (forward) ttggaagaattagatgtggt
PCR primer (reverse) TAACCACTGGGCCATTGTGT
Length of expected PCR product (bp) 417
Length of expected RE digestion product (bp) 205+212
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 72
Dosage of injected Cas9 mRNA (ng/ul) 1000
Dosage of injected gRNA (ng/ul) 300
Survival rate of F0 embryo @ 1dpf (%) 70
Method to estimate mutation efficiency Restriction enzyme digestionHpy1881
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO72.pdf
Mutation efficiency in F0 embryos (%) 30
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 30
Wild-type (WT) sequence GGTCGGTGTACGAGAGAAAACTGCAAGTACCTTCATCCACCGGCACATCTGAAGACGCAGCTGGAAATCAACGGACGCAACAACCTCATCCAGCAGA
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 GGTCGGTGTACGAGAGAAAACTGCAAGTACCTTCATCCA-----GACATCTGAAGACGCAGCTGGAAATCAACGGACGCAACAACCTCATCCAGCAGA (+1bp,-5bp)
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 20% GGTCGGTGTACGAGAGAAAACTGCAAGTACCTTCATCCA-----GACATCTGAAGACGCAGCTGGAAATCAACGGACGCAACAACCTCATCCAGCAGA (+1bp,-5bp)
F0-2
Allele name and predicted product 72a
72b
GGTCGGTGTACGAGAGAAAACTGCAAGTACCTTCATCCA-----GACATCTGAAGACGCAGCTGGAAATCAACGGACGCAACAACCTCATCCAGCAGA,Truncated protein(73aa)
突变创建时间 2015-06-18
填表时间 2013/12/16
修改时间 2016/12/29
PI/负责人 (工作单位)
Genotype Information: