Molecular characterization of ovary-specific gene Mrfem-1 and siRNA-mediated regulation on targeting Mrfem-1 in the giant freshwater prawn, Macrobrachium rosenbergii

•The Mrfem-1 cDNA from M. rosenbergii was isolated and characterized.•The Mrfem-1 mRNA were exclusively expressed in the ovary.•The Mrfem-1 transcripts were localized in different developmental stages oocytes.•Two siRNAs targeting Mrfem-1 were found and their effectiveness verified in vitro. Charact...

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Veröffentlicht in:Gene 2020-09, Vol.754, p.144891-144891, Article 144891
Hauptverfasser: Zhou, Ling-Xia, Liu, Xue, Ye, Bao-Qing, Liu, Yun, Tan, Shuang-Pei, Ma, Ke-Yi, Qiu, Gao-Feng
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container_title Gene
container_volume 754
creator Zhou, Ling-Xia
Liu, Xue
Ye, Bao-Qing
Liu, Yun
Tan, Shuang-Pei
Ma, Ke-Yi
Qiu, Gao-Feng
description •The Mrfem-1 cDNA from M. rosenbergii was isolated and characterized.•The Mrfem-1 mRNA were exclusively expressed in the ovary.•The Mrfem-1 transcripts were localized in different developmental stages oocytes.•Two siRNAs targeting Mrfem-1 were found and their effectiveness verified in vitro. Characterized by ankyrin repeat motifs, the feminization-1 (fem-1) gene plays an essential role in sex determination/differentiation in Caenorhabditis elegans. However, there are only a few reports on fem-1 in crustaceans. In this study, a fem-1 gene (Mrfem-1) was first isolated from the giant freshwater prawn Macrobrachium rosenbergii. The full-length cDNA of Mrfem-1 was 2607 bp long, containing an open reading frame encoding 615 amino acids, and presenting eight ankyrin repeats. The full-length cDNA has been submitted to GenBank with the accession no. MT160093. According to the RT-PCR results, Mrfem-1 was exclusively expressed in the ovary. The expression level of Mrfem-1 had increased with ovarian maturation and reached the highest peak at vitellogenic stage. In situ hybridization results showed that positive signals were concentrated in the cytoplasm of previtellogenic stage, and scattered in the cytoplasm and follicular cells at vitellogenic stage, suggesting that Mrfem-1 might be associated with ovarian maturation. Moreover, two effective siRNAs targeting Mrfem-1 were found and their effectiveness verified in vitro. These results on Mrfem-1 will help us to better understand the fem family and provide a new resource for subsequent investigations of siRNA-mediated regulation on ovarian development in M. rosenbergii.
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Characterized by ankyrin repeat motifs, the feminization-1 (fem-1) gene plays an essential role in sex determination/differentiation in Caenorhabditis elegans. However, there are only a few reports on fem-1 in crustaceans. In this study, a fem-1 gene (Mrfem-1) was first isolated from the giant freshwater prawn Macrobrachium rosenbergii. The full-length cDNA of Mrfem-1 was 2607 bp long, containing an open reading frame encoding 615 amino acids, and presenting eight ankyrin repeats. The full-length cDNA has been submitted to GenBank with the accession no. MT160093. According to the RT-PCR results, Mrfem-1 was exclusively expressed in the ovary. The expression level of Mrfem-1 had increased with ovarian maturation and reached the highest peak at vitellogenic stage. In situ hybridization results showed that positive signals were concentrated in the cytoplasm of previtellogenic stage, and scattered in the cytoplasm and follicular cells at vitellogenic stage, suggesting that Mrfem-1 might be associated with ovarian maturation. Moreover, two effective siRNAs targeting Mrfem-1 were found and their effectiveness verified in vitro. 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Characterized by ankyrin repeat motifs, the feminization-1 (fem-1) gene plays an essential role in sex determination/differentiation in Caenorhabditis elegans. However, there are only a few reports on fem-1 in crustaceans. In this study, a fem-1 gene (Mrfem-1) was first isolated from the giant freshwater prawn Macrobrachium rosenbergii. The full-length cDNA of Mrfem-1 was 2607 bp long, containing an open reading frame encoding 615 amino acids, and presenting eight ankyrin repeats. The full-length cDNA has been submitted to GenBank with the accession no. MT160093. According to the RT-PCR results, Mrfem-1 was exclusively expressed in the ovary. The expression level of Mrfem-1 had increased with ovarian maturation and reached the highest peak at vitellogenic stage. In situ hybridization results showed that positive signals were concentrated in the cytoplasm of previtellogenic stage, and scattered in the cytoplasm and follicular cells at vitellogenic stage, suggesting that Mrfem-1 might be associated with ovarian maturation. Moreover, two effective siRNAs targeting Mrfem-1 were found and their effectiveness verified in vitro. These results on Mrfem-1 will help us to better understand the fem family and provide a new resource for subsequent investigations of siRNA-mediated regulation on ovarian development in M. rosenbergii.</description><subject>Expression pattern</subject><subject>Feminization-1</subject><subject>Macrobrachium rosenbergii</subject><subject>siRNA</subject><issn>0378-1119</issn><issn>1879-0038</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><recordid>eNp9kU1r3DAQhkVJoZu0f6AnHXuoN_ry2oZcQuhHIJtCac9ClkdeLba0GWkT2v_T_1ktDjlmLgPDPPPxvoR85GzNGd9c7tcjBFgLJkpBqbbjb8iKt01XMSbbM7Jismkrznn3jpyntGcl6lqsyL9tnMAeJ4PU7gwamwH9X5N9DDQ6Gh8N_qnSAax33tLTErpFB3PFqQkDTf7n_XU1w-BNhoEijGXUAgeaDY6QfRhfEF-KO6CjNyFTh5B2T4VDekDzFD7TrbEY-3LEzh9nijFB6AFH79-Tt85MCT485wvy--uXXzffq7sf325vru8qK6XMVSvM0JUnYSO6fpD1UIMSzjnBG-WkdZ3pFAPbb1TXNK7fOAGsVgJ4UyvJZC8vyKdl7gHjwxFS1rNPFqbJBIjHpIXiinEpBC-tYmktJ6eE4PQB_VzU0pzpkyd6r09y6ZMnevGkQFcLBOWJRw-ok_UQbNEPwWY9RP8a_h8RG5f5</recordid><startdate>20200905</startdate><enddate>20200905</enddate><creator>Zhou, Ling-Xia</creator><creator>Liu, Xue</creator><creator>Ye, Bao-Qing</creator><creator>Liu, Yun</creator><creator>Tan, Shuang-Pei</creator><creator>Ma, Ke-Yi</creator><creator>Qiu, Gao-Feng</creator><general>Elsevier B.V</general><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-3834-3928</orcidid></search><sort><creationdate>20200905</creationdate><title>Molecular characterization of ovary-specific gene Mrfem-1 and siRNA-mediated regulation on targeting Mrfem-1 in the giant freshwater prawn, Macrobrachium rosenbergii</title><author>Zhou, Ling-Xia ; Liu, Xue ; Ye, Bao-Qing ; Liu, Yun ; Tan, Shuang-Pei ; Ma, Ke-Yi ; Qiu, Gao-Feng</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c333t-82ad9111e629bd35d5e42fff2174f3cf9a940ecb64977fb6f2e0542e1754303b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Expression pattern</topic><topic>Feminization-1</topic><topic>Macrobrachium rosenbergii</topic><topic>siRNA</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zhou, Ling-Xia</creatorcontrib><creatorcontrib>Liu, Xue</creatorcontrib><creatorcontrib>Ye, Bao-Qing</creatorcontrib><creatorcontrib>Liu, Yun</creatorcontrib><creatorcontrib>Tan, Shuang-Pei</creatorcontrib><creatorcontrib>Ma, Ke-Yi</creatorcontrib><creatorcontrib>Qiu, Gao-Feng</creatorcontrib><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Gene</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zhou, Ling-Xia</au><au>Liu, Xue</au><au>Ye, Bao-Qing</au><au>Liu, Yun</au><au>Tan, Shuang-Pei</au><au>Ma, Ke-Yi</au><au>Qiu, Gao-Feng</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Molecular characterization of ovary-specific gene Mrfem-1 and siRNA-mediated regulation on targeting Mrfem-1 in the giant freshwater prawn, Macrobrachium rosenbergii</atitle><jtitle>Gene</jtitle><date>2020-09-05</date><risdate>2020</risdate><volume>754</volume><spage>144891</spage><epage>144891</epage><pages>144891-144891</pages><artnum>144891</artnum><issn>0378-1119</issn><eissn>1879-0038</eissn><abstract>•The Mrfem-1 cDNA from M. rosenbergii was isolated and characterized.•The Mrfem-1 mRNA were exclusively expressed in the ovary.•The Mrfem-1 transcripts were localized in different developmental stages oocytes.•Two siRNAs targeting Mrfem-1 were found and their effectiveness verified in vitro. Characterized by ankyrin repeat motifs, the feminization-1 (fem-1) gene plays an essential role in sex determination/differentiation in Caenorhabditis elegans. However, there are only a few reports on fem-1 in crustaceans. In this study, a fem-1 gene (Mrfem-1) was first isolated from the giant freshwater prawn Macrobrachium rosenbergii. The full-length cDNA of Mrfem-1 was 2607 bp long, containing an open reading frame encoding 615 amino acids, and presenting eight ankyrin repeats. The full-length cDNA has been submitted to GenBank with the accession no. MT160093. According to the RT-PCR results, Mrfem-1 was exclusively expressed in the ovary. The expression level of Mrfem-1 had increased with ovarian maturation and reached the highest peak at vitellogenic stage. In situ hybridization results showed that positive signals were concentrated in the cytoplasm of previtellogenic stage, and scattered in the cytoplasm and follicular cells at vitellogenic stage, suggesting that Mrfem-1 might be associated with ovarian maturation. Moreover, two effective siRNAs targeting Mrfem-1 were found and their effectiveness verified in vitro. These results on Mrfem-1 will help us to better understand the fem family and provide a new resource for subsequent investigations of siRNA-mediated regulation on ovarian development in M. rosenbergii.</abstract><pub>Elsevier B.V</pub><doi>10.1016/j.gene.2020.144891</doi><tpages>1</tpages><orcidid>https://orcid.org/0000-0002-3834-3928</orcidid></addata></record>
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subjects Expression pattern
Feminization-1
Macrobrachium rosenbergii
siRNA
title Molecular characterization of ovary-specific gene Mrfem-1 and siRNA-mediated regulation on targeting Mrfem-1 in the giant freshwater prawn, Macrobrachium rosenbergii
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