Objective：This study aims to identify the 3′-UTR sequence of the ECSIT spliceosome in adult mice，and verify the effects of non-coding RNA on ECSIT expression. Methods：Using RACE technology to clone the mice ECSIT 3′-UTR non-coding region sequence，and further compare with the genomic database. Predict the possible binding microRNAs，and design siRNA against the full-length sequence of ECSIT. The expression levels of ECSIT in cells were determined by Western blot after using different microRNAs and siRNAs. Results：The results show that the 346 bp mouse ECSIT 3′-UTR was successfully identified and the sequence consistency rate with NCBI reached 99%. In cells，miR-7-5p and siRNA1，2 can interfere with the expression of ECSIT. Conclusion：These results shows that the 3′-UTR sequence of mouse ECSIT mRNA was successfully identified and this non-coding region can be used as a regulatory region of non-coding RNA，which can provide a scientific basis at the molecular level for further proving research of ECSIT in mouse growth and pathophysiological condition.