We present a novel method for treating bladder cancer with intravesically 
delivered small activating RNA (saRNA) in an orthotopic xenograft mouse bladder 
tumor model. The mouse model is established by urethral catheterization under 
inhaled general anesthetic. Chemical burn is then introduced to the bladder 
mucosa using intravesical silver nitrate solution to disrupt the bladder 
glycosaminoglycan layer and allows cells to attach. Following several washes 
with sterile water, human bladder cancer KU-7-luc2-GFP cells are instilled 
through the catheter into the bladder to dwell for 2 hours. Subsequent growth of 
bladder tumors is confirmed and monitored by in vivo bladder ultrasound and 
bioluminescent imaging. The tumors are then treated intravesically with saRNA 
formulated in lipid nanoparticles (LNPs). Tumor growth is monitored with 
ultrasound and bioluminescence. All steps of this procedure are demonstrated in 
the accompanying video.