Biomedical subjects

Wuming Gong

Publications and source records attributed to Wuming Gong.

8 recordsLinked to original sources

PubMed2006

Two novel transcripts encoding two Ankyrin repeat containing proteins have preponderant expression during the mouse spermatogenesis.

The clone 4921537P18 expressed preponderantly in mouse testis was identified by screening the Riken cDNA database, and two new full-length isoforms of this clone, which were named gsarp1 (Gonad Specific Ankyrin Repeat (ANK) Protein 1) and gsarp2, were found and isolated from mouse testis in the course of the research. Both of the GSARP1 and GSARP2 contain an ANK region circular composed by seven ANKs, and their structural feature is very similar to that of the IkappaB family proteins, while IkappaB proteins associate with the transcription factor NF-kappaB via their ANKs in the NF-kappaB pathway. We investigated the expression pattern at the mRNA level by Reverse transcription PCR. The gsarp1 has high expression level in mouse testis, while has low expression level in the ovary, and the gsarp2 is only expressed in mouse testis. The gsarp1 and gsarp2 begin to be detected at the early and later pachytene stage of meiosis separately, while both have high-expression level at the stage of MI and MII. The result of in situ hybridization reveals that the gsarp1 is primarily expressed in spermatocytes, while gsarp2 is expressed in spermatocytes and spermatids. In view of the structural feature and expression pattern of the GSARP1 and GSARP2, we speculate that they may play a certain role in a signal pathway of meiosis.

Amino Acid Sequence↗

PubMed2006

Integrated siRNA design based on surveying of features associated with high RNAi effectiveness.

BACKGROUND: Short interfering RNAs have allowed the development of clean and easily regulated methods for disruption of gene expression. However, while these methods continue to grow in popularity, designing effective siRNA experiments can be challenging. The various existing siRNA design guidelines suffer from two problems: they differ considerably from each other, and they produce high levels of false-positive predictions when tested on data of independent origins. RESULTS: Using a distinctly large set of siRNA efficacy data assembled from a vast diversity of origins (the siRecords data, containing records of 3,277 siRNA experiments targeting 1,518 genes, derived from 1,417 independent studies), we conducted extensive analyses of all known features that have been implicated in increasing RNAi effectiveness. A number of features having positive impacts on siRNA efficacy were identified. By performing quantitative analyses on cooperative effects among these features, then applying a disjunctive rule merging (DRM) algorithm, we developed a bundle of siRNA design rule sets with the false positive problem well curbed. A comparison with 15 online siRNA design tools indicated that some of the rule sets we developed surpassed all of these design tools commonly used in siRNA design practice in positive predictive values (PPVs). CONCLUSION: The availability of the large and diverse siRNA dataset from siRecords and the approach we describe in this report have allowed the development of highly effective and generally applicable siRNA design rule sets. Together with ever improving RNAi lab techniques, these design rule sets are expected to make siRNAs a more useful tool for molecular genetics, functional genomics, and drug discovery studies.

Algorithms↗

PubMed2006

siRecords: an extensive database of mammalian siRNAs with efficacy ratings.

UNLABELLED: Short interfering RNAs (siRNAs) have been gaining popularity as the gene knock-down tool of choice by many researchers because of the clean nature of their workings as well as the technical simplicity and cost efficiency in their applications. We have constructed siRecords, a database of siRNAs experimentally tested by researchers with consistent efficacy ratings. This database will help siRNA researchers develop more reliable siRNA design rules; in the mean time, siRecords will benefit experimental researchers directly by providing them with information about the siRNAs that have been experimentally tested against the genes of their interest. Currently, more than 4100 carefully annotated siRNA sequences obtained from more than 1200 published siRNA studies are hosted in siRecords. This database will continue to expand as more experimentally tested siRNAs are published. AVAILABILITY: The siRecords database can be accessed at http://siRecords.umn.edu/siRecords/

Abstracting and Indexing↗

PubMed2006

Zebrafish z-otu, a novel Otu and Tudor domain-containing gene, is expressed in early stages of oogenesis and embryogenesis.

Several studies have suggested that Otu domain had de-ubiquitinating activity and Tudor domain was important for the formation of germ cells. Here, we reported a novel zebrafish ovary-specific gene containing Otu and Tudor domain, z-otu, which was expressed at stages I-III oocytes and embryonic stages from zygotes to early blastula during embryonic cells maintained their totipotency. Therefore, z-otu might link the ubiquitin signaling pathway to early oogenesis and maintaining the totipotency of embryonic cell.

Amino Acid Sequence↗

PubMed2006

PSM2, a novel protein similar to MCAF2, is involved in the mouse embryonic and adult male gonad development.

By screening RIKEN database, we obtained a mouse ovary and uterus derived clone (RIKEN clone ID: 5031403I04). Multiple tissues expression analysis revealed that the clone was specifically expressed in ovary and to a higher extent in testis. It is expressed in early mouse embryo, especially in the embryonic gonad from 11.5 dpc (days post coitus). Furthermore, it is also expressed continuously from newborn testis to adult. Using testis sections in situ hybridization, we found the mRNA was localized to spermatogonia, round spermatids and mainly to spermatocytes. We cloned the cDNA from mouse testis. The gene consists of 10 exons spanning approximately 48 kb on mouse chromosome 16. The cDNA encodes a putative nuclear protein of 319 amino acids containing a coiled-coil motif. The deduced protein has high similarity with human MCAF2 (MBD1-containing chromatin-associated factor 2), so we termed it as PSM2 (protein similar to MCAF2) in this article. We therefore hypothesized that PSM2 might interact with some important partners by the conserved domain and be involved in the transcription modulation during gonad development.

Amino Acid Sequence↗

PubMed2006

The identification and characterization of a testis-specific cDNA during spermatogenesis.

Using bioinformatics and experimental validation, we obtained a cDNA (named srsf) which was exclusively expressed in the mouse testes. RT-PCR analysis showed that srsf mRNA was not expressed in the gonad during the sex determination period or during embryogenesis. In developing mouse testis, srsf expression was first detected on post-natal day 10, reached its highest level on day 23, and then reduced to and remained at a moderate level throughout adulthood. In situ hybridization analysis demonstrated that srsf mRNA was expressed in pachytene spermatocytes and round spermatids in the testes. The predicted protein contains one RNA-binding domain (RBD) and a serine-arginine rich domain (RS), which are characterized by some splicing factors of SR family members. These findings indicate that srsf may play a role during spermatogenesis.

Amino Acid Sequence↗

PubMed2005

Cloning and characterization of full length of a novel zebrafish gene Zsrg abundantly expressed in the germline stem cells.

Using the digital differential display program of the National Center for Biotechnology Information, we identified a contig of expression sequence tags (ESTs) (Accession No. BM316936), which came from zebrafish ovary and testis libraries. The full-length cDNA of this transcript was cloned and further confirmed by polymerase chain reaction and sequencing. The full-length cDNA of the novel gene is 807bp and encodes a novel protein of 187 amino acids, which shares no significant homology with any other known proteins. Characterization of genomic sequences of the gene revealed that it spans 6kb on the linkage group 3 and is composed of five exons and four introns. RT-PCR analysis showed that it was expressed in mature oocytes and one-cell stage, and persisted until 24h of development. RT-PCR also revealed that it is expressed in gonad and kidney, with the highest level of expression in the testis. The expression sites of the novel gene in adult gonad were further localized by in situ hybridization to oogonia and growing oocytes in ovary and to spermatogonia, spermatocytes but not to spermatids in testis. Based on its abundance in testis and the germline stem cell-spermatogonia and oogonia, we hypothesize that it may function as a testicular development and gametogenesis related gene that plays important roles in spermatogenesis, and named it Zsrg (zebrafish testis spermatogenesis related gene, Zsrg).

Amino Acid Sequence↗

PubMed2005

Identification of a new transcript specifically expressed in mouse spermatocytes: mmrp2.

In an effort to examine the molecular basis of gametogenesis, we screened Riken cDNA database and the clone 4930481F22 that is expressed preponderantly in mouse testis was identified. In the course of the research, a new isoform of 4930481F22 clone was found, isolated from mouse testis and sequenced. It only lacks the 7th exon of 4930481F22 transcript. The new isoform only has 837 bp and encodes a putative 28.4 kDa protein. We investigated the expression pattern at the mRNA level by RT-PCR and in situ hybridization in testis. The new isoform was only expressed in the gonad, where it began to be detected at day 8 after birth. In situ hybridization proved that the new isoform mostly expressed in spermatocytes. The structure of the predicted protein and the expression pattern of the mRNA suggest that the new isoform could have an important role in meiosis. We temporarily named it mmrp 2 (Mouse Meiosis Related Protein 2).

Amino Acid Sequence↗