Antisense drug discovery: Can cell-free screens speed the process?

被引:14
作者
Branch, AD [1 ]
机构
[1] CUNY Mt Sinai Sch Med, Dept Med, Div Liver Dis, New York, NY 10029 USA
来源
ANTISENSE & NUCLEIC ACID DRUG DEVELOPMENT | 1998年 / 8卷 / 03期
关键词
D O I
10.1089/oli.1.1998.8.249
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Many conditions must be satisfied for an antisense drug to function. It must colocalize with its target RNA at a sufficient concentration for a bimolecular reaction to occur, and it must have a structure that favors association with its target. In addition, if the antisense compound is to form Watson-Crick bonds with the target RNA, it must be complementary to sites that are amenable to binding. Unfortunately, the peculiarities that cause certain sites to be especially vulnerable to antisense compounds are undefined, as discussed previously (Branch, 1998), Because vulnerable target sites have no common properties allowing them to be identified by sequence analysis, most target sites and their antisense counterparts are found through a trial and error process in which oligomers-each complementary to a different site in the target RNA-are tested individually to find the one with the greatest specificity and lowest inhibitory concentration (IC,,), However, testing antisense molecules one at a time can be a taxing process, and there is great interest in developing cell-free screening methods that can reduce the number of compounds that must be tested in cells and in whole animals. These cell-free screens are designed to generate short lists of target sites that include the ideal site-the site most vulnerable to antisense ablation in vivo. They are based on the unproven assumption that ideal sites have distinctive properties, such as susceptibility to RNase H-mediated cleavage, that allow them to be detected in cell-free assays. This is a review of data emerging from studies using RNase H-based screens and a summary of the challenges confronting these and any similar methods that use naked RNAs as surrogates for intracellular RNAs, It is not yet clear if cell-free screening methods will be effective.
引用
收藏
页码:249 / 254
页数:6
相关论文
共 13 条
[1]   A good antisense molecule is hard to find [J].
Branch, AD .
TRENDS IN BIOCHEMICAL SCIENCES, 1998, 23 (02) :45-50
[2]   Efficient hammerhead ribozyme and antisense RNA targeting in a slow ribosome Escherichia coli mutant [J].
Chen, H ;
Ferbeyre, G ;
Cedergren, R .
NATURE BIOTECHNOLOGY, 1997, 15 (05) :432-435
[3]  
Cioffi CL, 1997, MOL PHARMACOL, V51, P383
[4]   The role of innovation in drug development [J].
Drews, J ;
Ryser, S .
NATURE BIOTECHNOLOGY, 1997, 15 (13) :1318-1319
[5]   Evaluation of retinal toxicity and efficacy of anti-cytomegalovirus and anti-herpes simplex virus antiviral phosphorothioate oligonucleotides ISIS 2922 and ISIS 4015 [J].
FloresAguilar, M ;
Besen, G ;
Vuong, C ;
Tatebayashi, M ;
Munguia, D ;
Gangan, P ;
Wiley, CA ;
Freeman, WR .
JOURNAL OF INFECTIOUS DISEASES, 1997, 175 (06) :1308-1316
[6]   Potent antisense oligonucleotides to the human multidrug resistance-1 mRNA are rationally selected by mapping RNA-accessible sites with oligonucleotide libraries [J].
Ho, SP ;
Britton, DHO ;
Stone, BA ;
Behrens, DL ;
Leffet, LM ;
Hobbs, FW ;
Miller, JA ;
Trainor, GL .
NUCLEIC ACIDS RESEARCH, 1996, 24 (10) :1901-1907
[7]   Mapping of RNA accessible sites for antisense experiments with oligonucleotide libraries [J].
Ho, SP ;
Bao, YJ ;
Lesher, T ;
Malhotra, R ;
Ma, LY ;
Fluharty, SJ ;
Sakai, RR .
NATURE BIOTECHNOLOGY, 1998, 16 (01) :59-63
[8]   Selecting effective antisense reagents on combinatorial oligonucleotide arrays [J].
Milner, N ;
Mir, KU ;
Southern, EM .
NATURE BIOTECHNOLOGY, 1997, 15 (06) :537-541
[9]   Antitumor activity of a phosphorothioate antisense oligodeoxynucleotide targeted against C-raf kinase [J].
Monia, BP ;
Johnston, JF ;
Geiger, T ;
Muller, M ;
Fabbro, D .
NATURE MEDICINE, 1996, 2 (06) :668-675
[10]  
MUCCIOLI C, 1998, RETR OPP INFC CHIC