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Nucleic Acids Research, 1994, Vol. 22, No. 12 2242-2248
© 1994

RNA

Reduced ß2-microglobulin mRNA levels in transgenic mice expressing a designed hammerhead ribozyme

Sten Larsson, Graham Hotchkiss, Michael Andäng, Tommy Nyholm, José Inzunza, Irma Jansson and Lars Ährlund-Richter*

Unit for Molecular Genetics, Center for Biotechnology, Karolinska Institute Novum, 141 57 Huddinge, Sweden

*To whom correspondence should be addressed

Received April 8, 1994. Revised May 25, 1994. Accepted May 25, 1994.

We have generated three artificial hammerhead ribozymes, denoted ‘Rz-b’, ‘Rz-c’ and ‘Rz-d’, with different specificities for exon II of the mouse beta-2-microglobulin (ß2M) mRNA. In this study we tested for ribozyme mediated reduction of ß2M mRNA in a cell line and in transgenic mice. Transfections of either of the Rz-b, Rz-c or Rz-d plasmids into a mouse cell-line (NIH/3T3) revealed reductions of ß2M mRNA substrate in each case. Ribozyme expression in individual transfected clones was accompanied with an up to 80% reduction of ß2M mRNA levels. Rz-c was selected for a transgenic study. Seven Rz-c transgenic founder animals were identified from which three ribozyme expressing families were established and analysed. Expression of the ribozyme transgene was tested for and detected in lung, kidney and spleen. Expression was accompanied with reduction of the ß2M mRNA levels of heterozygous (Rz +/–) animals compared to non-transgenic litter mates. The effect was most pronounced in lung with more than 90% ß2M mRNA reduction in individual mice. In summary, expression of our ribozymes in a cell free system, in a cell-line and in transgenic mice were all accompanied with reductions of ß2M mRNA levels.


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