Objective To design the ribozymes which may cleave the mRNA of human TIMP-1. Establishing plasmids transcripting the ribozymes to produce abundant ribozymes. Methods The TIMP-1 mRNA was taken as the target RNA. Ribozymes were designed according to the hammerhead structure described by Symons. To establish clones of TIMP-1 ribozymes use P 1.5. Then obtain abundant ribozymes by transcripting. Results Three ribozymes targeting the nt 123, nt 299 and nt 353 on TIMP-1 mRNA were designed. The ribozyme gene has been synthesized, plasmids cloned and abundant ribozymes obtained. Conclusion Computer assisted design is indispensable in studying ribozyme. The establishment of plasmids transcripting ribozymes is the key step of studying their activity.