• Molecules and Cells
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• 제23권3호
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• pp.316-322
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• 2007

Zinc finger transcription factor genes are a significant fraction of the genes in the vertebrate genome. Here we report the isolation and characterization of a human zinc finger-containing gene, ZNF435, from a fetal brain cDNA library. ZNF435 cDNA is 1290 base pairs in length and contains an open reading frame encoding 349 amino acids with four C2H2-type zinc fingers at its carboxyl terminus and a SCAN motif at its amino terminus. RT-PCR results showed that ZNF435 was expressed in all tested tissues. A ZNF435-GFP fusion protein was located in the nucleus and the four zinc fingers acted as nuclear localization signals (NLSs). ZNF435 was found to be capable of homo-association, and this effect was independent of its zinc fingers. Furthermore, ZNF435 proved to be a transcription repressor as its overexpression in AD293 cells inhibited the transcriptional activities of AP-1.

• BMB Reports
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• 제40권4호
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• pp.517-524
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• 2007

There were many different families of zinc finger proteins that contained multiple cysteine and/or histidine residues and used zinc to stabilize their folds. The classical C2H2 zinc finger proteins were the founding members of this superfamily and were among the most abundant proteins in eukaryotic genomes. C2H2 proteins typically contained several C2H2 fingers that made tandem contacts along the DNA. Here we reported a novel C2H2 type zinc finger gene, ZNF438, which encoded 828 amino acids that formed five zinc finger domains. Bioinformatics analysis revealed that the ZNF438 was mapped to human chromosome 10p11.2 and shared 62% identity with rat and mouse homologues. RT-PCR analysis indicated that it was ubiquitously expressed in 18 human adult tissues. With immunofluorescence assay, it was shown that the exogenous Flag-tagged ZNF438 was located in nucleus of COS-7 cells. To further explore the function of ZNF438, we examined the transcriptional activity of ZNF438 protein by transfecting recombinant pM-ZNF438 into mammalian cells. The subsequent analysis based on the duel luciferase assay system showed that ZNF438 was a transcriptional repressor.

• Parasites, Hosts and Diseases
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• 제40권1호
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• pp.59-64
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• 2002

A Cryptosporidium parvum sporozoite and oocyst λgt11 cDNA library was screened with a hyperimmune rabbit serum that was developed against insoluble fragments of ultrasonicated oocysts. A clone named Cp22.4.1 encoding a protein of 231 amino acids with 4 zinc-finger domains characterized by a Cys-X2-Cys-X4-His-X4-Cys motif was isolated and characterized. There was a complete match between the sequencing data of the coding region of Cp22.4.1 and the corresponding gene at chromosomal level. Cloning in a pBAD-TOPO-TA expression vector permitted to evaluate the antigenicty of the recombinant His-tagged antigen. This antigen was recognized by 2 out of 5 sera from Cruptosporidium immune calves and not by sera from parasite naive animals.

• Plant Biotechnology Reports
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• 제5권1호
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• pp.9-17
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• 2011

With the development of next-generation sequencing technology, ever-expanding databases of genetic information from various organisms are available to researchers. However, our ability to study the biological meaning of genetic information and to apply our genetic knowledge to produce genetically modified crops and animals is limited, largely due to the lack of molecular tools to manipulate genomes. Recently, targeted cleavage of the genome using engineered DNA scissors called zinc finger nucleases (ZFNs) has successfully supported the precise manipulation of genetic information in various cells, animals, and plants. In this review, we will discuss the development and applications of ZFN technology for genome engineering and highlight recent reports on its use in plants.

• Mass Spectrometry Letters
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• 제6권1호
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• pp.7-12
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• 2015

In the present study, we proposed a simple ESI-MS model for determining $Zn^{2+}$ binding (or dissociation) constants for zinc finger peptides (ZFPs) with a unique ${\beta}{\beta}{\alpha}$ fold consensus. The ionization efficiency (response) factors for this model, i.e., ${\alpha}$ and ${\beta}$, could be determined for ZiCo ZFP with a known $Zn^{2+}$ binding constant. We could determine the binding constants for other ZFPs assuming those with a ${\beta}{\beta}{\alpha}$ consensus conformation have the same ${\alpha}/{\beta}$ response ratio. In general, the ZPF dissociation constants exhibited $K_d$ values of $10^{-7}{\sim}10^{-9}M$, while $K_d$ values for a negative control non-specific $Zn^{2+}$ peptides were high, e.g., $5.5{\times}10^{-6}M$ and $4.3{\times}10^{-4}M$ for BBA1 and melittin, respectively.

• KSBB Journal
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• 제25권3호
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• pp.215-222
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• 2010

Monogenic disease의 치료를 위한 하나의 전략으로 viral vector를 이용한 gene therapy에 비해 독성이 적은 gene targeting 기술을 이용하기 위한 연구가 진행되고 있다. 이러한 연구의 주된 관점은 자연적인 HR의 낮은 효율을 개선하기 위한 DSB 유도 방법으로, 선택성을 높일 수 있는 긴 염기서열의 인식이 가능한 artificial endonuclease의 개발이다. 본 글에서는 이러한 artificial endonuclease 중, 가장 많이 연구 되고 있는 homing endonuclease와 zinc finger nuclease를 간략히 소개하였다. 전자와 후자 모두, 인식 서열에 대한 일정 수준의 tolerance (인식 서열 일부가 특이적이지 않아 다른 염기로 구성된 경우)가 존재하여, 일정한 비율로 다른 target을 절단할 수 있는 가능성이 존재한다. 이러한 점은, meganucleases를 치료 목적으로 이용할 때 세포 독성을 나타내는 근본원인 중 하나이다. 두 종 모두 이러한 특성을 가짐에도 불구하고, 완전한 비자연적인 후자보다는 전자의 경우가 보다 효과적이며 낮은 세포독성을 보이는 것으로 보고되고 있다. 물론 실험 조건이나 적용되는 세포 종류, 인위적인 단백질의 발현 정도에 따라 세포 독성유무 또는 정도에 차이가 나타남이 확인되고 있다. 이러한 사실들에 근거할 때, gene targeting을 유도하기 위한 artificial endonuclease의 서열 특이성을 증대시키는 것이 가장 중요하나, 그 외 여러 인자들에 대한 복합적인 연구 역시 필요함을 보여준다. 현재까지 실제 치료제로 쓰인 예는 없지만, 시험관내에서 보이는 결과와 모델 개체에서 이루어진 표적화정도, 관련된 단백질 치료제들이 지닌 잠재성을 비교할 때 매우 큰 가능성을 지니고 있음은 충분히 확인할 수 있다.

• BMB Reports
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• 제53권3호
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• pp.160-165
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• 2020

The root meristem of Arabidopsis thaliana is protected by the root cap, the size of which is tightly regulated by the balance between the formative cell divisions and the dispersal of the outermost cells. We isolated an enhancer-tagged dominant mutant displaying the short and twisted root by the overexpression of ZINC-FINGER OF ARABIDOPSIS THALIANA1 (ZAT1) encoding an EAR motif-containing zinc-finger protein. The growth inhibition by ZAT1 was shared by ZAT4 and ZAT9, the ZAT1 homologues. The ZAT1 promoter was specifically active in the outermost cells of the root cap, in which ZAT1-GFP was localized when expressed by the ZAT1 promoter. The outermost cell-specific expression pattern of ZAT1 was not altered in the sombrero (smb) or smb bearskin1 (brn1) brn2 accumulating additional root-cap layers. In contrast, ZAT4-GFP and ZAT9-GFP fusion proteins were distributed to the inner root-cap cells in addition to the outermost cells where ZAT4 and ZAT9 promoters were active. Overexpression of ZAT1 induced the ectopic expression of PUTATIVE ASPARTIC PROTEASE3 involved in the programmed cell death. The EAR motif was essential for the growth inhibition by ZAT1. These results suggest that the three related ZATs might regulate the maturation of the outermost cells of the root cap.

• Genomics & Informatics
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• 제4권4호
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• pp.147-160
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• 2006

GATA transcription factors are widespread eukaryotic regulators whose DNA-binding domain is a class IV zinc finger motif in the form $CX_{2}CX_{17-20}CX_{2}C$followed by a basic region. In fungi, they act as transcriptional activators or repressors in several different processes, ranging from nitrogen source utilization to mating-type switching. Using an in-house bioinformatics portal system, we surveyed 50 fungal and 9 out-group genomes and identified 396 putative fungal GATA transcription factors. The proportion of GATA transcription factors within a genome varied among taxonomic lineages. Subsequent analyses of phylogenetic relationships among the fungal GATA transcription factors, as well as a study of their domain architecture and gene structure, demonstrated high degrees of conservation in type IVa and type IVb zinc finger motifs and the existence of distinctive clusters at least at the level of subphylum. The SFH1 subgroup with a 20-residue loop was newly identified, in addition to six well-defined subgroups in the subphylum Pezizomycotina. Furthermore, a novel GATA motif with a 2f-residue loop ($CX_{2}CX_{21}CX_{2}C$, designated 'zinc finger type IVc') was discovered within the phylum Basidiomycota. Our results suggest that fungal GATA factors might have undergone multiple distinct modes of evolution resulting in diversified cellular modulation in fungi.

• Asian-Australasian Journal of Animal Sciences
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• 제29권10호
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• pp.1500-1507
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• 2016

Myostatin (MSTN) can negatively regulate the growth and development of skeletal muscle, and natural mutations can cause "double-muscling" trait in animals. In order to block the inhibiting effect of MSTN on muscle growth, we transferred zinc-finger nucleases (ZFN) which targeted sheep MSTN gene into cultured fibroblasts. Gene targeted colonies were isolated from transfected fibroblasts by serial dilution culture and screened by sequencing. Two colonies were identified with mono-allele mutation and one colony with bi-allelic deletion. Further, we introduced the MSTN-ZFN mRNA into sheep embryos by microinjection. Thirteen of thirty-seven parthenogenetic embryos were targeted by ZFN, with the efficiency of 35%. Our work established the technical foundation for generation of MSTN gene editing sheep by somatic cloning and microinjection ZFN into embryos.

• Journal of Plant Biotechnology
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• 제34권3호
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• pp.167-172
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• 2007

A tomato zinc-finger protein gene, LeZFP1, encoding the Cys2/His2-type zinc-finger transcription factor was searched from cDNA microarray analysis of gene expression following induction of the overexpressed tomato transgenic plants showing resistance for pathogen and abiotic stresses. The full-length cDNA of LeZFP1 encoded a protein of 261 amino acid residues. Analysis of the deduced amino acid sequence of LeZFP1 revealed that it shares high sequence identity with pepper CAZFP1 (81% identity). We found that single copy of LeZFP1 gene is present in the tomato genome through southern blot analysis. The LeZFP1 transcripts were constitutively expressed in the tomato mature and young leaves, but were detectable weakly in the flower, stem and root. The LeZFP1 transcripts were significantly reduced in treated leaf tissues with NaCl and mannitol. The LeZFP1 gene was induced by oxidative stress especially. Our results indicated that LeZFP1 may play a role function involved in oxidative stress signaling pathways.