OBJECTIVE: To explore the genetic basis for a child featuring short stature and postaxial polydactyly. METHODS: A child who presented at Ningbo Women & Children's Hospital in May 2021 due to the"discovery of growth retardation for more than two years" was selected as the subject. Peripheral blood samples of the child and his parents were collected for the extraction of genomic DNA. Whole exome sequencing was carried out for the child, and candidate variant was verified by Sanger sequencing of his family members. RESULTS: The child was found to harbor a heterozygous c.3670C>T (p.Q1224) variant of the GLI2 gene, which may lead to premature termination of protein translation. The variant was not detected in either parent. CONCLUSION The child was diagnosed with Culler-Jones syndrome. The c.3670C>T (p.Q1224*) variant of the GLI2 gene probably underlay the disease in this child.
Child ; Female ; Humans ; Fingers ; Mutation ; Nuclear Proteins/genetics* ; Polydactyly/genetics* ; Toes ; Zinc Finger Protein Gli2/genetics*

OBJECTIVETo compare binding activity of different zinc finger domain of human Kaiso with methylated CpG.

METHODSpGEX constructs with different human Kaiso domain were generated and then corresponding fusion proteins were induced and purified. Electrophoretic mobility shift assays were applied to evaluate the binding activity of fusion proteins with methylated CpG.

RESULTSThe purified GST-KaisoZF fusion protein (without the POZ protein binding domain) could bind with methylated CpG probe specifically, but not for three or two zinc fingers without flanking domains.

CONCLUSIONHuman zinc finger protein Kaiso could bind with methylated CpG specifically, only in the assistance of the neighboring flank sequence of the zinc finger domain.

Base Sequence ; CpG Islands ; DNA Methylation ; Humans ; Transcription Factors ; genetics ; Zinc Fingers ; genetics

Zinc finger nuclease (ZFN), which is a chimeric fusion structure between a Cys2-His2 zinc-finger protein (ZFP) and the cleavage domain of Fok I endonuclease, can be used to introduce targeted double-stranded breaks (DSBs). ZFN-mediated cleavage leads to mutations when double-stranded breaks are repaired by homologous recombination (HR) or nonhomologous end joining (NHEJ). In recent years, ZFNs are widely used in the fields of genetic research. In this review, the methodology and technical advantages of ZFNs were briefly discussed.
Animals ; Deoxyribonucleases, Type II Site-Specific ; chemistry ; genetics ; metabolism ; Humans ; Zinc Fingers

As the ubiquitous nucleic acids recognizing motif, Zinc finger protein play important role in regulation of gene expression. The study of recognization specific will greatly facilitate understanding the delicate interaction of Zinc finger protein and DNA. By the choice of expression vector, the induction and culture conditions, the DNA binding domain of Zif268 was expressed in Escherichia coli partly solubly. The gel mobility shift assay shows that purified DNA binding domain can bind its natural target sequence specifically, which indicates the DNA binding domain remains its DNA binding activity in Escherichia coli. The functional expression of DNA binding domain of Zif268 will greatly facilitate the development of in vivo genetic selection assay for the study of Zinc fingers-DNA interaction.
DNA-Binding Proteins ; genetics ; Electrophoretic Mobility Shift Assay ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; Zinc Fingers ; genetics

CTCF as a multivalent eukaryotic transcription factor plays a diverse range of roles in regulation of various genes through the binding of its 11 zinc fingers to CTCF consensus sites or various proteins. CTCF is involved in multiple aspects of epigenetic regulation including regulation of chromatin remodeling and genomic imprinting. Deregulation of these processes result in a group of diseases are characterized by growth, development, and neurological dysfunction. This paper reviews recent researches that highlight the links between CTCF, epigenetics and diseases.
CCCTC-Binding Factor ; Disease ; genetics ; Epigenomics ; methods ; Humans ; Repressor Proteins ; genetics ; Transcription Factors ; genetics ; Zinc Fingers ; genetics

Precise and effective modification of complex genomes at the predicted loci has long been an important goal for scientists. However, conventional techniques for manipulating genomes in diverse organisms and cells have lagged behind the rapid advance in genomic studies. Such genome engineering tools have featured low efficiency and off-targeting. The newly developed custom-designed nucleases, zinc finger nucleases (ZFN), transcription activator-like effector nucleases (TALEN) and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system have conferred genome modification with ease of customization, flexibility and high efficiency, which may impact biological research and studies on pathogenesis of human diseases. These novel techniques can edit the genomic DNA with high efficiency and specificity in a rich variety of organisms and cell types including the induced pluripotent stem cells (iPSCs), which has conferred them with the potential for revealing the pathogenesis and treatment of many human diseases. This review has briefly introduced the mechanisms of ZFN, TALENs and CRISPR/Cas9 system, and compared the efficiency and specificity of such approaches. In addition, the application of ZFN, TALENs and CRISPR/Cas9 mediated genome modification for human disease modeling and gene therapy was also discussed.
Base Sequence ; CRISPR-Cas Systems ; genetics ; Genetic Therapy ; methods ; Humans ; Transcription Activator-Like Effector Nucleases ; genetics ; Zinc Fingers ; genetics

Transcription factor is a key trans-acting factor to mediate stress response by regulating gene expression. Plants have developed a series of mechanisms to modulate development, stress response, signaling and disease resistance at transcription level. DNA binding with one finger (DOF), containing one C₂-C₂ zinc finger domain, is a special plant transcription factor. Specifically, the conserved domain at N-terminus of DOF has multiple functions, including interacting with DNA and protein, which could be involved in plant development and stress response. Although many DOF family genes are characterized in plant stress response, it is not clear if DOF genes have functions in cereal plants. In the present paper, the role of DOF family genes on cereal plants were discussed based on a comprehensive phylogenetic relationship analysis, expression profiles in different tissues and various environmental conditions. The results obtained here will provide an important reference for further understanding the mechanism of gramineous crops in stress resistance.
DNA-Binding Proteins ; metabolism ; Gene Expression Regulation, Plant ; Phylogeny ; Plant Proteins ; metabolism ; Plants ; genetics ; Transcription Factors ; metabolism ; Zinc Fingers

Breeding of robust industrial Saccharomyces cerevisiae strains with high ethanol tolerance is of great significance for efficient fuel ethanol production. Zinc finger proteins play important roles in gene transcription and translation, and exerting control on the regulation of multiple genes. The sequence and localization of the zinc finger motif can be designed and engineered, and the artificial zinc finger protein can be used to regulate celluar metabolism. Stress tolerance of microbial strains is related to multiple genes. Therefore, it is possible to use artificially-designed zinc finger proteins to breed stress tolerant strains. In this study, a library containing artificial zinc finger protein encoding genes was transformed into the model yeast strain S288c. A recombinant strain named M01 with improved ethanol tolerance was obtained. The plasmid in M01 was isolated, and then transformed into the industrial yeast strain Sc4126. Ethanol tolerance of the recombinant strain of Sc4126 were significantly improved. When high gravity ethanol fermentation using 250 g/L glucose was performed, comparing with the wild-type strain, fermentation time of the recombinant strain was decreased by 24 h and the final ethanol concentration was enhanced by 6.3%. The results of this study demonstrate that artificial zinc finger proteins are able to exert control on stress tolerance of yeast strains, and these results provide basis to construct robust industrial yeast strains for efficient ethanol fermentation.
Adaptation, Physiological ; drug effects ; Drug Resistance, Fungal ; genetics ; Ethanol ; pharmacology ; Fungal Proteins ; genetics ; metabolism ; Industrial Microbiology ; Mutation ; genetics ; Peptide Library ; Saccharomyces cerevisiae ; genetics ; growth & development ; Zinc Fingers

In plants, proteins with A20/AN1 zinc finger domain are involved in stress responses, named as "Stress Associated Protein" (SAP) gene family. Based on Expressed Sequence Tag (EST) sequences information in Badila Saccharum officinarum mature related cDNA library, we cloned an SAP gene from sugarcane full length cDNA library, named ShSAP1 (GenBank: Accession No. HM991960). To characterize ShSAP1, we analyzed its genome structure and expression pattern. Southern blot analysis showed ShSAP1 was present as one or two copy in the genome of Badila. Comparison of ShSAP1 1 008 bp full length cDNA with a genomic frangment (2 241 bp) generated by PCR amplification and sequencing, revealed the presence of two introns (202 bp and 1 052 bp) located in the 5'UTR region. Semiquantitative RT-PCR analysis found ShSAP1 expressed in leaves, roots and stalk in mature sugarcane. Compared with immature stems, ShSAP1 expressed higher in mature stalk. ShSAP1 was induced by different types of treatments, such as salt (200 mmol/L NaCl), drought (10% PEG 6 000), GA3 (200 mg/L), ABA (100 micromol/L) and ET (1 mmol/L) during sugarcane seedling stage. These results indicated that ShSAP1 may function in sugarcane maturation and abiotic stress response processes.
Amino Acid Sequence ; Cloning, Molecular ; Gene Expression Regulation, Plant ; Molecular Sequence Data ; Plant Proteins ; genetics ; Saccharum ; genetics ; metabolism ; Stress, Physiological ; genetics ; Zinc Fingers ; genetics

OBJECTIVETo investigate the association of Apa I polymorphism in vitamin D receptor (VDR) gene with bone mass in men.

METHODSThe VDR Apa I genotype was determined by PCR-restriction fragment length polymorphism (RFLP) in 388 unrelated healthy men aged 46-80 years of Han nationality in Shanghai city. Bone mineral density (BMD) and bone mineral content (BMC) at lumber spine 1-4 (L1-4) and at any sites of proximal femur including to femoral neck (Neck), trochanter (Troch) and Ward's striangle (Ward's) were measured by duel-energy X-ray absorptiometry.

RESULTSFrequencies distribution of VDR Apa I genotype were aa for 48.1%, Aa for 44.2% and AA 7.7%. The allele frequencies of Apa I polymorphism were in Hardy-Weinberg equilibrium. No significant association was found between Apa I genotype and BMD or BMC in group of all population or in subgroup of men below 60 years. In men above 60 years, the significant association was found between VDR Apa I genotype and BMD or BMC at L1-4, Neck and Ward's (P < 0.05, P < 0.01) and compared with Aa and aa genotype, AA genotype had significantly higher mean BMD and BMC at L1-4, Neck and Ward's (P < 0.05, P < 0.01). But Apa I genotype is not associated with BMD and BMC at Troch.

CONCLUSIONSApa I polymorphism is associated with bone mass in men above 60 years, and AA genotype has higher bone mass. Apa I polymorphism in VDR gene possibly influence loss of trabecular and cortical bone mass in old men.

Age Factors ; Aged ; Aged, 80 and over ; Alleles ; Bone Density ; Humans ; Male ; Middle Aged ; Phenotype ; Polymorphism, Restriction Fragment Length ; Receptors, Calcitriol ; genetics ; Transcription Factors ; genetics ; Zinc Fingers ; genetics