RNA transcription system using novel ribozyme

Details RNA transcription system using novel ribozyme RNA transcription system using novel ribozyme

1993-05-03

1996-03-19

Stone, Jacqueline M.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing compound containing saccharide radical

435 6, 4351723, 4352401, 4352402, 4352523, 4353201, 536 231, 536 232, 536 245, 514 44, C12N 922, C12N 1574, C12N 1582, C12N 1585

Patent

active

055003575

ABSTRACT:
The invention provides a recombinant plasmid containing a sequence encoding any genes inserted between 5' and 3' self-cleavage ribozymes. The recombinant plasmid can be amplified in vivo as well as in vitro while growing the host cell. When obtaining RNA transcripts of the inserted sequence, the recombinant plasmid does not require a restriction enzyme digestion step (run-off transcription) since cis-acting ribozymes perform self-catalyzed cleavage at 5' and 3' sides of the inserted sequence once it is transcribed. In this specific example, the trans-acting RNA enzyme sequence is inserted between 5' and 3' cleavage ribozymes. However, the trans-acting ribozyme sequence in the recombinant plasmid can be replaceable with any other sequence (e.g., antisense RNA, RNAs of HIV-1, HDV and other RNA viruses etc.). This construct is especially useful since each unit, consisting of 5' processing ribozyme, inserted sequence, and 3' processing ribozyme, can be connected in tandem. By so doing, ribozymes targeted to various sites can initially be transcribed as a long RNA chain which subsequently undergoes cleavage to produce independent trans-acting ribozymes, each possessing a specific target site.

REFERENCES:
Taira et al. (1990), Protein Engineering, vol. 3, No. 8, pp. 733-737.
Hutchins et al. (1986), Nucleic Acid Res., vol. 14, pp. 3627-3640.
Gerlach et al., Virology, vol. 151, pp. 172-185.
Cotten et al. (1989), EMBO J., vol. 8, pp. 3861-3866.
Diener et al. (1989), Proc. Natl. Acad. Sci., vol. 86, pp. 9370-9374.
Dzianott et al. (1989), Proc. Natl. Acad. Sci., vol. 86, pp. 4823-4827.

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