国家水生生物种质资源库国家斑马鱼资源中心 (China Zebrafish Resource Center)

ZKO number		ZKO13
Recent progress of this line		F2 individuals have been indentified
Gene name		cenpe
Ensembl ID		ENSDARG00000063385
Zebrafish strain		AB
Cas9 target site 1 sequence		sense strand GGCATCTGAGAGCTCGGAGCcgg
Cas9 target site 2 sequence
Position in the gene		exon 2
Restriction enzyme (RE)		Sac I(GAGCT\|C)
PCR primer (forward)		TACTATTTTATGTTCCTGTTATGG
PCR primer (reverse)		GCTCTACTACTTGGATTGCC
Length of expected PCR product (bp)		401
Length of expected RE digestion product (bp)		175+102+124
Plasmids used for making Cas9 mRNA		pXT7-hCAS9
Plasmids?used?for?making?gRNA		pMD19T-gRNA
Primers?used?for?making?gRNA		Pr-ZKO 13
Dosage of injected Cas9 mRNA (ng/ul)		300
Dosage of injected gRNA (ng/ul)		30
Survival rate of F0 embryo @ 1dpf (%)		95
Method to estimate mutation efficiency		Restriction enzyme digestionRE digestion (Sac I)
Stage of the embryos collected for mutation detection		http://www.zfish.cn/upload/file/ZKOPDF/ZKO13.pdf
Mutation efficiency in F0 embryos (%)		30
Picture?from?F₀?embryos ? (gel?picture?or?sequencing?picture)		/upload/2014-09-18/635466521277999948.png 13
Mutation type		Indel
Germline transmission efficiency in adult F₀		35%(6/17)
Wild-type (WT) sequence		GGCATCTGAGAGCTCGGAGC
Germline mutations in selected F0 (By screening F1 embryos)	F0-1
Germline mutations in selected F0 (By screening F1 embryos)	F0-2
Mosaicism in selected F0 (By screening F1 adults)	F0-1	GGCATCTGA-------GAGC(-7bp)
Mosaicism in selected F0 (By screening F1 adults)	F0-2
Allele name and predicted product	13a 13b	GGCATCTGA-------GAGC; Truncated protein(98aa)
Allele name and predicted product	13a 13b
突变创建时间		2014-04-10
填表时间		2013/12/18
修改时间		2016/8/8
PI/负责人 (工作单位)

Genotype Information: