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PCR-Based Construction of Long Hammerhead Ribozymes

  • Martin Zillmann
  • Gregory Robinson
Part of the Methods in Molecular Biology™ book series (MIMB, volume 74)

Abstract

With the advent of antisense technology, there has been much interest in the use of long RNAs expressed in vivo to inhibit the expression of target genes. More recently, there have been numerous reports that the incorporation of either hairpin or hammerhead ribozyme motifs (catalytic antisense) into such RNAs increases their effectiveness (1, 2, 3). This section will describe a generally applicable, simple, PCR-based method to construct catalytic antisense RNA containing the hammerhead catalytic core that will cleave the target of interest at a GUC sequence. All discussion will focus on the hammerhead motif, although, in principle, the hairpin motif could be incorporated instead.

Keywords

Microcentrifuge Tube Hammerhead Ribozyme Hairpin Motif Final Insert Incomplete Denaturation 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Humana Press Inc. 1997

Authors and Affiliations

  • Martin Zillmann
    • 1
  • Gregory Robinson
    • 1
  1. 1.HybridonWorcester

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