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Conference Paper: Functional study of a Novel Acrosome-specific Gene (AEP1/VAD1.3) using Conditional Gene Knock Out Approach

TitleFunctional study of a Novel Acrosome-specific Gene (AEP1/VAD1.3) using Conditional Gene Knock Out Approach
Authors
Issue Date2006
PublisherHong Kong Academy of Medicine.
Citation
The 4th International Huaxia Congress of Endocrinology (IHCE-4), Hong Kong, 15–18 December 2006. In Hong Kong Medical Journal, 2006, v. 12 n. 6 suppl. 4, p. 110, abstract no. P87 How to Cite?
AbstractObjective: AEP1/VAD1.3, a novel acrosome gene, was first identified from a retinol treated Vitamin A deficiency (VAD) rat model with synchronized spermatogenesis. AEP1 was expressed specifically in the testes from Day 25 when spermatids form. AEP1 immunoreactivity was localized at the acrosome region of the rat, human, monkey and porcine spermatids and spermatozoa, suggesting that AEP1 may play a significant role on acrosome formation. We hypothesized that AEP1 deficiency may cause acrosomal defect in mouse sperm leading to infertility. Methods: To elucidate the role of AEP1 during spermatogenesis, a Cre-LoxP conditional gene knock out approach will be used to generate AEP1 deficient mice for functional studies. Results: Conditional target vector was constructed by inserting the LoxP sites flanking the exons 2 and 3 of the AEP1 gene. A neo cassette flanked by Frt sites and a splice-receptor conjugated EGFP gene for in vitro selection of recombinant embryonic stem (ES) cells and Loxed AEP1 allele in sperm, respectively, were inserted downstream of the exons 3. In vitro recombinant occurred when the targeting vector was transformed into E. coli harboring Cre or Flp recombinases targeting for LoxP and Frt sites of the targeting vector, respectively. The conditional targeting construct was used to electrophorate 129/SvEv embryonic stem (ES) cells to generate conditional KO mice. Conclusion: The conditional knockout targeting vector for AEP1 gene was generated and characterized. Generation of AEP1 conditional knockout animal may allow us to understand the role of AEP1 on acrosome formation and infertility in human. (This work was supported in part by a RGC grant HKU7537/05M to KFL.)
Persistent Identifierhttp://hdl.handle.net/10722/113576
ISSN
2023 Impact Factor: 3.1
2023 SCImago Journal Rankings: 0.261

 

DC FieldValueLanguage
dc.contributor.authorZuo, Yen_HK
dc.contributor.authorTam, YTen_HK
dc.contributor.authorYeung, WSBen_HK
dc.contributor.authorLee, CKFen_HK
dc.date.accessioned2010-09-26T04:21:56Z-
dc.date.available2010-09-26T04:21:56Z-
dc.date.issued2006en_HK
dc.identifier.citationThe 4th International Huaxia Congress of Endocrinology (IHCE-4), Hong Kong, 15–18 December 2006. In Hong Kong Medical Journal, 2006, v. 12 n. 6 suppl. 4, p. 110, abstract no. P87-
dc.identifier.issn1024-2708-
dc.identifier.urihttp://hdl.handle.net/10722/113576-
dc.description.abstractObjective: AEP1/VAD1.3, a novel acrosome gene, was first identified from a retinol treated Vitamin A deficiency (VAD) rat model with synchronized spermatogenesis. AEP1 was expressed specifically in the testes from Day 25 when spermatids form. AEP1 immunoreactivity was localized at the acrosome region of the rat, human, monkey and porcine spermatids and spermatozoa, suggesting that AEP1 may play a significant role on acrosome formation. We hypothesized that AEP1 deficiency may cause acrosomal defect in mouse sperm leading to infertility. Methods: To elucidate the role of AEP1 during spermatogenesis, a Cre-LoxP conditional gene knock out approach will be used to generate AEP1 deficient mice for functional studies. Results: Conditional target vector was constructed by inserting the LoxP sites flanking the exons 2 and 3 of the AEP1 gene. A neo cassette flanked by Frt sites and a splice-receptor conjugated EGFP gene for in vitro selection of recombinant embryonic stem (ES) cells and Loxed AEP1 allele in sperm, respectively, were inserted downstream of the exons 3. In vitro recombinant occurred when the targeting vector was transformed into E. coli harboring Cre or Flp recombinases targeting for LoxP and Frt sites of the targeting vector, respectively. The conditional targeting construct was used to electrophorate 129/SvEv embryonic stem (ES) cells to generate conditional KO mice. Conclusion: The conditional knockout targeting vector for AEP1 gene was generated and characterized. Generation of AEP1 conditional knockout animal may allow us to understand the role of AEP1 on acrosome formation and infertility in human. (This work was supported in part by a RGC grant HKU7537/05M to KFL.)-
dc.languageengen_HK
dc.publisherHong Kong Academy of Medicine.-
dc.relation.ispartofHong Kong Medical Journalen_HK
dc.titleFunctional study of a Novel Acrosome-specific Gene (AEP1/VAD1.3) using Conditional Gene Knock Out Approachen_HK
dc.typeConference_Paperen_HK
dc.identifier.emailZuo, Y: ho494060@hkusua.hku.hken_HK
dc.identifier.emailTam, YT: h0202957@hkusua.hku.hken_HK
dc.identifier.emailYeung, WSB: wsbyeung@hkucc.hku.hken_HK
dc.identifier.emailLee, CKF: ckflee@hkucc.hku.hken_HK
dc.identifier.authorityYeung, WSB=rp00331en_HK
dc.identifier.authorityLee, CKF=rp00458en_HK
dc.identifier.hkuros134621en_HK
dc.identifier.volume12-
dc.identifier.issue6 suppl. 4-
dc.identifier.spage110, abstract no. P87-
dc.identifier.epage110, abstract no. P87-
dc.identifier.issnl1024-2708-

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