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The present invention provides a catalyst composition, comprising : an organic boron salt and a bisimine thioester rare earth complex having a structure of formula (I), with the molar ratio of the organic boron salt to the bisimine thioester rare earth complex having a structure of formula (I) being (0.5-2.0) : 1. The present invention also provides a method for preparing the catalyst composition, comprising the following steps : mixing and reacting the organic boron salt, the bisimine thioester rare earth complex having a structure of formula (I) of claim 1, and an organic solvent, so as to obtain the catalyst composition for preparing a crystalline 3, 4-polyisoprene. The catalyst composition for preparing a crystalline 3, 4-polyisoprene provided by the present invention has a high selectivity, and can result in a high isotactic crystalline 3, 4-polyisoprene with a higher content.