Genetic analysis of a family with congenital heart defects caused by chromosome 8p23.1 deletion / 中华医学遗传学杂志

Objective@#To explore the genetic basis for a family affected with congenital heart defects.@*Methods@#G-banding karyotyping, chromosomal microarray analysis (CMA) and multiplex ligation-dependent probe amplification (MLPA) were carried out to detect copy number variants in a patient with left ventricular noncompaction (LVNC) and his fetus.@*Results@#G-banding karyotyping showed the patient was 45, XY, rob(15; 21)(q10; q10)[36]/46, XY[64], while the fetus had an normal karyotype. CMA revealed that both had arr[hg19]8p23.1(11 232 919-11 935 465)×1. MLPA showed both had deletion of all exons of the GATA4 gene.@*Conclusion@#The LVNC of the patient and the ventricular septal defect(VSD) of his fetus may result from the same 8p23.1 deletion, for which GATA4 is probably the key gene.

Establishment and verification of a mouse model of gene H435Y mutation / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a mouse model of H435Y mutation of gene using CRISPR/Cas9- mediated gene targeting.</p><p><b>METHODS</b>The single-stranded guide RNA (sgRNA) specific to the H435Y loci of gene was designed based on the sequence of gene. After activity assessment, the active sgRNA and Cas9 were transcribed into RNA and microinjected along with the donor DNA fragment with point mutations into fertilized mouse eggs. The microinjected eggs were transferred into pseudopregnant mice to obtain the F0 generation mice with the target gene mutation confirmed by PCR and gene sequencing. gene mutations in the offsprings of the F0 generation mice were analyzed.</p><p><b>RESULTS</b>Gene sequencing confirmed the successful establishment of mouse models carrying H435Y mutation of gene in 4 of the F0 generation mice. The positive F0 generation mice were crossed with wild-type C57BL/6J mice to obtain the F1 generation mice, and PCR confirmed the presence of H435Y mutations of gene in 6 of the F1 mice. Then F2 generation mice were obtained by F1 generation matting with each other. PCR showed that H435Y mutation of gene in F2 mice was found, indicating the mousemodel of gene mutation in H435Y was established and propagated successfully.</p><p><b>CONCLUSIONS</b>We successfully established gene H435Y mutant mouse models using CRISPR/Cas9 technique.</p>

GATA4 gene mutation in congenital heart disease / 国际儿科学杂志

GATA4 gene mutations are common in children with congenital heart disease.More than 120 mutations have been found in ventricular septal defect,atrial septal defects and the tetralogy of fallot.But the relationship between genotype and clinical phenotype has not yet been elucidated.This article summarizes the published germline and somatic mutations of GATA4 gene in human congenital heart disease,and provides insight into the phenotypic spectrum of GATA4 gene mutation.

Knockout gata4 gene and establish ment of a zebrafish model of congenital heart disease by TALEN / 中国比较医学杂志

Objective To establish a gata4 gene knockout zebrafish model of congenital heart disease, and construct transcription activator-like effector nuclease ( TALEN) vectors targeting gata4 gene.Method We construct TALEN vectors targeting zabrafish gata4 gene using unit assembly method and the in vitro-transcribed TALEN mRNAs were microinjected into one-cell stage zebrafish embryos.The efficiency of TALEN was identified by injected embryos, and mutations of zebrafish were screened and confirmed the different types through PCR and enzyme digestion.Results We successfully constructed correct targeting vectors by enzyme digestion and sequencing, and the gene knockout efficiency was 35.18%.We screened the mutant zebrafish and confirmed different types of gata4 gene mutations.Conclusions A gata4 knockout zebrafish model is successfully established, it can provide a good animal model for further research of congenital heart diseases.

Mutation analysis of GATA4 gene in Han Chinese patients with atrioventricular septal defect / 临床儿科杂志

Objective To analyze the mutations of GATA4 gene in Han Chinese patients with atrioventricular septal defect(AVSD)and investigate the association between GATA4 gene and pathogenesis. Methods Ninety-four Han Chinese patients with AVSD were recruited,including 23 patients with Down syndrome and 71 patients without. One hundred healthy age-matched Han children were used as the control. Blood samples were drawn. Encoding region and flanking introns of GATA4 gene were amplified using polymerase chain reaction. The mutations were detected by DNA fragment sequences analysis. Results Three novel missense mutations(c.106C ＞ G,p.P36A;c.259C ＞ T, p.P87S;c.504C ＞ A,p.D168E)of the GATA4 gene were identified in three patients with complete AVSD without Down syndrome,and a fourth novel missense mutation(c.1079A ＞ G,p.E360G)was noted in a patient with complete AVSD and Down syndrome. A polymorphism of the GATA4 gene(c.G99T,p.A33A)from six patients was detected. Conclusions The GATA4 gene might be involved in the etiology of AVSD by functional changes resulting from gene mutation. The low incidence of GATA4 gene mutations in patients with AVSD with or without Down syndrome might suggest that AVSD is a polygenetic disorder.

Genetic Reveal Study of NKX2.5,TBX5 and GATA4 Genes in Patients With Tetralogy of Fallot / 中国循环杂志

Objective:To investigate mRNA expression of cardiac related genes of NKX2.5,TBX5 and GATA4 in patients with tetralogy of fallot(TOF). Methods:A total of 10 TOF patients(TOF group)from 4 months to 8 years with the mean age of 3.5 years were recruited in our hospital from June to December 2005.The patients were diagnosed by typical clinical manifestation and cardiac color echocardiogram, the diagnosis was confirmed by cardiac surgery.6 non-congenital heart disease children were selected as Control group, and they were from 4 months to 9 years with the mean age of 3.8 years.The myocardial total RNA was extracted,the related cDNA was obtained by RT-PCR.The product cDNA was amplified with fluorescent quantitative PCR in order to compare the differences of NKX2.5,TBX5,GATA4 and GAPDH mRNA expression between TOF group and Control group. Results:NKX2.5 mRNA expression in TOF group was statistically decreased than that in Control group,while there were no statistical changes found in TBX5 and GATA4 mRNA expression between TOF group and Control group. Conclusion:The mRNA expression of NKX2.5,TBX5 and GATA4 were found in myocardium development.TOF was possibly related to decreased NKX2.5 mRNA expression.