title: Lessons learned from vivo-morpholinos: How to avoid vivo-morpholino toxicity.
abstract: Vivo-morpholinos are a promising tool for gene silencing. These oligonucleotide analogs transiently silence genes by blocking either translation or pre-mRNA splicing. Little to no toxicity has been reported for vivo-morpholino treatment. However, in a recent study conducted in our lab, treatment of mice with vivo-morpholinos resulted in high mortality rates. We hypothesized that the deaths were the result of oligonucleotide hybridization, causing an increased cationic charge associated with the dendrimer delivery moiety of the vivo-morpholino. The cationic charge increased blood clot formation in whole blood treated with vivo-morpholinos, suggesting that clotting could have caused cardiac arrest in the deceased mice. Therefore, we investigate the mechanism by which some vivo-morpholinos increase mortality rates and propose techniques to alleviate vivo-morpholino toxicity.
authors.affiliation: Children's Nutrition Research Center, Baylor College of Medicine, Houston, TX; Biology of Physical Activity Laboratory, Dept. of Health & Kinesiology, Texas A&M University, College Station, TX.
authors.forename: David P
authors.initials: DP
authors.lastname: Ferguson
authors.affiliation: Protein Chemistry Laboratory, Dept. of Biochemistry/Biophysics, Texas A&M University, College Station, TX.
authors.forename: Lawrence J
authors.initials: LJ
authors.lastname: Dangott
authors.affiliation: Biology of Physical Activity Laboratory, Dept. of Health & Kinesiology, Texas A&M University, College Station, TX.
authors.forename: J Timothy
authors.initials: JT
authors.lastname: Lightfoot
mesh_terms: D000818:Animals; D017317:Annexin A6 / Q000235:genetics; D001777:Blood Coagulation / Q000187:drug effects; D002135:Calcium-Binding Proteins / Q000235:genetics; D002155:Calsequestrin; D004305:Dose-Response Relationship, Drug; D020868:Gene Silencing; D007267:Injections; D008297:Male; D008810:Mice, Inbred C57BL; D060172:Morpholinos / Q000008:administration & dosage / Q000633:toxicity*; D009026:Mortality; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage / Q000633:toxicity*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D017317:Annexin A6; D002135:Calcium-Binding Proteins; D002155:Calsequestrin; C503130:Casq1 protein, mouse; D060172:Morpholinos; D016376:Oligonucleotides, Antisense
keywords: annexin A6; blood clot formation; calsequestrin 1; vivo-morpholino