Splicing and stability of intron in the expression retroviral vector with human clotting factor IX

Yong Na Xing, Da Ru Lu, Xiaobo Gao, Xin Fang Qiu, Jing Lun Xue

Research output: Contribution to journalArticle

Abstract

To study the role of intron in the expression of hFIX, retroviral vectors with intron containing hFIX were constructed. It is fundamental for the intron study whether the intron constructed in retroviral vector can be steadily transferred into target cell. First, we constructed two forward-orientation retroviral vectors: GlNaC-i-IX contains the exogenous intron from IL-2, and GlNaC-i′ -IX contains the truncated intron I from hFIX gene, covering the splicing donor and acceptor sequences. RT-PCR result indicated that intron in the forward-orientation retroviral vector was spliced after packaging in PA317. Then, reverse-orientation retroviral vectors GlNaC-i′ -IXR and GlNaPAIXi ′ BAM were constructed, in which the reverse and complimentary sequences of hfIX gene with intron appeared in retroviral RNA. RT-PCR assay combined with ELISA test indicated that intron was retained after packaging and hFIX gene with intron constructed in the reverse-orientation retroviral vector can be transduced intact and expressed hFIX. at a high level in vitro.

Original languageEnglish (US)
Pages (from-to)476-477
Number of pages2
JournalActa Genetica Sinica
Volume25
Issue number6
StatePublished - Dec 1 1998
Externally publishedYes

Fingerprint

Factor IX
Blood Coagulation Factors
Introns
Product Packaging
Genes
Polymerase Chain Reaction
Interleukin-2
Enzyme-Linked Immunosorbent Assay
RNA

Keywords

  • Clotting factor IX
  • Endogenous intron
  • Exogenous intron
  • Retroviral vector
  • Splicing

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Splicing and stability of intron in the expression retroviral vector with human clotting factor IX. / Xing, Yong Na; Lu, Da Ru; Gao, Xiaobo; Qiu, Xin Fang; Xue, Jing Lun.

In: Acta Genetica Sinica, Vol. 25, No. 6, 01.12.1998, p. 476-477.

Research output: Contribution to journalArticle

Xing, Yong Na ; Lu, Da Ru ; Gao, Xiaobo ; Qiu, Xin Fang ; Xue, Jing Lun. / Splicing and stability of intron in the expression retroviral vector with human clotting factor IX. In: Acta Genetica Sinica. 1998 ; Vol. 25, No. 6. pp. 476-477.
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AU - Xue, Jing Lun

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AB - To study the role of intron in the expression of hFIX, retroviral vectors with intron containing hFIX were constructed. It is fundamental for the intron study whether the intron constructed in retroviral vector can be steadily transferred into target cell. First, we constructed two forward-orientation retroviral vectors: GlNaC-i-IX contains the exogenous intron from IL-2, and GlNaC-i′ -IX contains the truncated intron I from hFIX gene, covering the splicing donor and acceptor sequences. RT-PCR result indicated that intron in the forward-orientation retroviral vector was spliced after packaging in PA317. Then, reverse-orientation retroviral vectors GlNaC-i′ -IXR and GlNaPAIXi ′ BAM were constructed, in which the reverse and complimentary sequences of hfIX gene with intron appeared in retroviral RNA. RT-PCR assay combined with ELISA test indicated that intron was retained after packaging and hFIX gene with intron constructed in the reverse-orientation retroviral vector can be transduced intact and expressed hFIX. at a high level in vitro.

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