In vitro synthesized SV40 cRNA is trans‐spliced after microinjection into the nuclei of mammalian cells

FEBS Letters

Graessmann, A.¹  Graessmann, M.¹  Eul, J.¹ 

[1] Institut für Molekularbiologie und Biochemie der Freien Universität Berlin, Arnimallee 22, 14195 Berlin, Germany

关键词: SV40 T1-antigen;    Pre-mRNA synthesis;    Microinjection;    Trans-splicing;    Translation;   

DOI  :  10.1016/0014-5793(96)00957-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We present for the first time experimental evidence that in vitro synthesized RNA (cRNA) is trans-spliced after microinjection into the nuclei of mammalian tissue culture cells. The template used for cRNA synthesis was the early SV40 BstXI/BamHI DNA fragment. This DNA fragment encodes exclusively for the second T-antigen exon and contains the intact small t-antigen intron. To generate the corresponding mRNA (t1-mRNA) by trans-splicing, the cells utilize a 5′ cryptic splice site located within the second T-antigen exon of one cRNA molecule which is spliced to the small t-antigen 3′ splice site of another cRNA molecule. Formation of the T1-mRNA by trans-splicing was confirmed by RT-PCR analysis and DNA sequencing. Efficient trans-splicing required that competitive small t-antigen cis-splicing be inhibited by deletion of the small t-antigen 5′ splice site. The T1-mRNA was not generated when the cryptic 5′ splice site was mutated.

【 授权许可】

Unknown