Premium
mRNA Structural Modelling Predicts Variability in Efficacy of Post‐transcriptional Gene Silencing Between Species
Author(s) -
Trujillo Alexandria J,
Sullivan Jack M
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.818.8
Subject(s) - ribozyme , hammerhead ribozyme , gene silencing , messenger rna , rna , gene , computational biology , biology , genetics
Post‐transcriptional Gene Silencing ( PTGS ) agents, such as hammerhead ribozymes, face unique barriers when designed against an mRNA target. Hammerhead ribozymes( hhRzs ) are able to cleave ‘NUH’ (N= any nucleotide H= U, A, C) sites in target mRNA. Along with sequence specificity, the NUH site must be located in a region of the mRNA that is accessible after secondary and tertiary structural folding. Our lab has developed a streamlined RNA Drug Discovery ( RDD ) platform and designed hhRzs that target the 266 and 1362 regions in Rhodopsin( RHO ) mRNA, to develop a treatment for Autosomal Dominant Retinitis Pigmentosa, a disease that causes progressive vision loss. Bioinformatic analysis using mRNA structure prediction programs MFold, SFold, and RNA structure was conducted on fourteen species representatives to determine if the 266 and 1362 regions have similar accessibility for our ribozymes. The analysis shows that even in cases of sequence homology and close evolutionary relationships, access to the target region varies among species compared to human RHO . This suggests that during RDD efficacy and toxicity studies of PTGS agents in vitro and in vivo should be conducted against human RHO and transgenic animals expressing human RHO for the most accurate predictions. Support or Funding Information This work was supported majorly by NIH/NEI R01 grant EY013433 (JMS), and partially by the Department of Veterans Affairs (VA), Veterans Health Administration, Office of Research and Development (Biomedical Laboratory Research and Development) (VA Merit Grant 1I01BX000669 (JMS)), and partially by an Unrestricted Grant from Research to Prevent Blindness to the Department of Ophthalmology at the University at Buffalo‐SUNY, and also by the Interdisciplinary Science and Engineering Partnership (NSF 1102998 PI: Joseph Gardella (AJT)). The study was conducted in its entirety at the Veterans Administration Western New York Healthcare System (Buffalo, NY). Contents do not represent the views of the Department of Veterans Affairs or the United Stated Government.