Cloning and expression characterization of the chicken Piwil1 gene

Chen, Rong; Chang, Guobin; Dai, Aiqin; Ma, Teng; Zhai, Fei; Xia, Mingxiu; Liu, Lu; Li, Jianchao; Hua, Dengke; Chen, Guohong

doi:10.1007/s11033-013-2831-9

Cited by 8 publications

(5 citation statements)

References 31 publications

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“…In many other mammalian species, PIWIL1 expression is highly restricted to the testes in both developing and mature animals [27,45,46,57]. The pattern of PIWIL1 expression reported here is consistent with that reported in other species using either an N-terminal or C-terminal-specific antibody.…”

Section: Discussionsupporting

confidence: 89%

cDNA Cloning of Feline PIWIL1 and Evaluation of Expression in the Testis of the Domestic Cat

Stalker

Backx

Tscherner

et al. 2023

IJMS

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The PIWI clade of Argonaute proteins is essential for spermatogenesis in all species examined to date. This protein family binds specific classes of small non-coding RNAs known as PIWI-interacting RNAs (piRNAs) which together form piRNA-induced silencing complexes (piRISCs) that are recruited to specific RNA targets through sequence complementarity. These complexes facilitate gene silencing through endonuclease activity and guided recruitment of epigenetic silencing factors. PIWI proteins and piRNAs have been found to play multiple roles in the testis including the maintenance of genomic integrity through transposon silencing and facilitating the turnover of coding RNAs during spermatogenesis. In the present study, we report the first characterization of PIWIL1 in the male domestic cat, a mammalian system predicted to express four PIWI family members. Multiple transcript variants of PIWIL1 were cloned from feline testes cDNA. One isoform shows high homology to PIWIL1 from other mammals, however, the other has characteristics of a “slicer null” isoform, lacking the domain required for endonuclease activity. Expression of PIWIL1 in the male cat appears limited to the testis and correlates with sexual maturity. RNA-immunoprecipitation revealed that feline PIWIL1 binds small RNAs with an average size of 29 nt. Together, these data suggest that the domestic cat has two PIWIL1 isoforms expressed in the mature testis, at least one of which interacts with piRNAs.

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Section: Discussionsupporting

confidence: 89%

cDNA Cloning of Feline PIWIL1 and Evaluation of Expression in the Testis of the Domestic Cat

Stalker

Backx

Tscherner

et al. 2023

IJMS

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“…In avian species, the biological functions of PIWI family members remain unclear. In recent studies, we characterized the chicken PIWI members CIWI and CILI (Kim et al, ), and revealed the complete genomic structure of the CIWI gene, including a 161‐bp fragment of the 5'‐UTR and 660 bp of the 3'‐UTR followed by a poly(A) tail (Chen et al, ).…”

Section: Discussionmentioning

confidence: 99%

“…PCR products of the correct size were cloned into the pGEM T easy vector (Promega, Madison, WI). A 4‐kb genomic fragment from −3,839 to +161 bp, including the predicted transcription start site, was cloned, but the +1 to +161‐bp region includes the 5'‐UTR and does not include the ATG start codon (Chen et al, ). The eGFP‐coding sequence and polyadenylation (poly‐A) tail were inserted into the cloned vectors containing the CIWI promoter using restriction enzymes Spe I and Nde I.…”

Section: Methodsmentioning

confidence: 99%

The CCAAT element in the CIWI promoter regulates transcriptional initiation in chicken primordial germ cells

Sohn

Lee

Kim

et al. 2014

Molecular Reproduction Devel

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The P-element-induced wimpy testis (PIWI) protein, which associates with small non-coding RNAs, is responsible for maintaining the integrity of germ cells and stem cells. Thus, transcriptional regulation of PIWI is critical for its effective functional modulation. In this study, we identified the promoter region of the PIWI homolog in chicken (CIWI), and investigated the transcriptional regulatory elements that control expression of CIWI in chicken primordial germ cells (PGCs). We constructed a vector that included the enhanced green fluorescent protein (eGFP) gene controlled by the 4-kb CIWI promoter. The vector was expressed in chicken PGCs, but not in chicken embryonic fibroblasts. Based on promoter deletion and fragmentation assays, we found that a 252-bp fragment of the CIWI promoter (-577 to -326 bp) was crucial for CIWI expression in PGCs. A CCAAT transcriptional regulatory element (-498 to -494 bp) was detected in the proximal region from the transcription initiation site of CIWI, and mutational analysis confirmed that this element regulates transcriptional initiation in chicken PGCs. Interestingly, the regions flanking the CCAAT element, which are positioned differently in HIWI (human), Miwi (mouse), and CIWI orthologs, were highly conserved. In addition, we predicted that specificity protein 1 (SP1) motifs modulate the transcriptional initiation of CIWI by binding to the 5'-flanking regions of the CCAAT box. Overall, 252 bp of the CIWI promoter possessing the transcriptional regulatory element CCAAT is crucial for regulating CIWI gene expression in chicken PGCs. This promoter may be applicable for the regulation of CIWI expression during germ-cell development.

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“…there are three types of PIWI proteins in mouse which are associated with piRNAs and expressed in male mouse germline and they are essential for fertility (Carmell et al, 2007). In chicken there is not enough studies about biological function of PIWI proteins however in a study PIWI like1 was detected in chicken and it was suggested that it has more expression in gonad and pays roles in myosis I (Chen et al, 2013). VASA protein belongs to the Dead-box helicase (DDX) family (Lasko & Ashburner, 1988).…”

Section: Important Genes Regulating Pgcs Specificationmentioning

confidence: 99%

In vitro culture of reptile PGCS to preserve endangered species

Golkar‐Narenji

Dzięgiel

Kempisty

et al. 2023

Cell Biology International

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Primordial germ cells (PGCs), are the source of gametes in vertebrates. There are similarities in the development of PGCs of reptiles with avian and mammalian species PGCs development. PGCs culture has been performed for avian and mammalian species but there is no report for reptilian PGCs culture. In vitro culture of PGCs is needed to produce transgenic animals, preservation of endangered animals and for studies on cell behaviour and research on fertility. Reptiles are traded as exotic pets and a source of food and they are valuable for their skin and they are useful as model for medical research. Transgenic reptile has been suggested to be useful for pet industry and medical research. In this research different aspects of PGCs development was compared in three main classes of vertebrates including mammalian, avian and reptilian species. It is proposed that a discussion on similarities between reptilian PGCs development with avian and mammalian species helps to find clues for studies of reptilian PGCs development details and finding an efficient protocol for in vitro culture of reptilian PG.

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Cloning and expression characterization of the chicken Piwil1 gene

Cited by 8 publications

References 31 publications

cDNA Cloning of Feline PIWIL1 and Evaluation of Expression in the Testis of the Domestic Cat

cDNA Cloning of Feline PIWIL1 and Evaluation of Expression in the Testis of the Domestic Cat

The CCAAT element in the CIWI promoter regulates transcriptional initiation in chicken primordial germ cells

In vitro culture of reptile PGCS to preserve endangered species

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