NDLI: Generating in vitro transcripts with homogenous 3′ ends using trans-acting antigenomic delta ribozyme

Content Provider	PubMed Central
Author	Wichłacz, Agnieszka Łęgiewicz, Michał Ciesiołka, Jerzy
Copyright Year	2004
Abstract	In most in vitro run-off transcription reactions with T7 RNA polymerase, transcripts with heterogeneous ends are commonly obtained. Towards the goal of finding a simple and effective procedure for correct processing of their 3′ ends we propose the use of trans-acting antigenomic delta ribozyme. We demonstrate that the extension of nascent transcripts with only seven nucleotides complementary to the ribozyme’s recognition site, and subsequently, the removal of those nucleotides with the ribozyme acting in trans, is an efficient procedure for generating transcripts with homogenous 3′ ends. This approach was tested on two model RNA molecules: an in vitro transcript of yeast tRNAPhe and a delta ribozyme, which processed itself during transcription. The proposed procedure is a simple alternative to the use of ribozymes as cis-cleaving autocatalytic cassettes attached to transcript 3′ ends. As there is little possibility that the required additional stretch, only seven nucleotides long, enters into stable interactions with other parts of the transcripts, it can be cleaved off with high efficacy.
Related Links	http://dx.doi.org/10.1093/nar/gnh037
Ending Page	39
Starting Page	39
File Format	PDF
ISSN	13624962
e-ISSN	13624962
Journal	Nucleic Acids Research
Issue Number	3
Volume Number	32
Language	English
Publisher	Oxford University Press
Publisher Date	2004-02-13
Access Restriction	Open
Rights Holder	Oxford University Press
Subject Keyword	Genetics Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Genetics

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