Abstract:
:We report the use of an engineered ribozyme to produce a circular human exon in vitro. Specifically, we have designed a derivative of a yeast self-splicing group II intron that is able to catalyze the formation of a circular exon encoding the first kringle domain (K1) of the human tissue plasminogen activator protein. We show that the circular K1 exon is formed with high fidelity in vitro. Furthermore, the system is designed such that the circular exon that is produced consists entirely of human exon sequence. Thus, our results demonstrate that all yeast exon sequences are dispensable for group II intron catalyzed inverse splicing. This is the first demonstration that an engineered ribozyme can be used to create a circular exon containing only human sequences, linked together at a precise desired ligation point. We expect these results to be generalizable, so that similar ribozymes can be designed to precisely create circular derivatives of any nucleotide sequence.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Mikheeva S,Hakim-Zargar M,Carlson D,Jarrell Kdoi
10.1093/nar/25.24.5085subject
Has Abstractpub_date
1997-12-15 00:00:00pages
5085-94issue
24eissn
0305-1048issn
1362-4962pii
gka808journal_volume
25pub_type
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