By Yogesh S. Sanghvi, Mark Andrade, Rantjit R. Deshmukh, Lars Holmberg (auth.), Gunther Hartmann, Stefan Endres (eds.)
In the prior few years, antisense method has moved from in vitro experiences to in vivo experiences and primary human trials. whereas the fundamental inspiration of antisense know-how is easy, the methodological difficulties linked to its use are a number of and intricate. Antisense- dependent equipment have confirmed to be a box of study the place cautious consciousness to experimental protocols and acceptable controls is important.
The Manual of Antisense Methodology emphasizes the applying of antisense oligonucleotides, and is a consultant for the identity of antisense and non-antisense results in several experimental settings. The paintings is geared up into 3 sections: antisense software in vitro, antisense software in vivo (animal versions) and at last, medical antisense reports. the place in any respect attainable, the equipment are defined in enough aspect to permit copy of a given test.
The Manual of Antisense Methodology could be of curiosity to researchers in immunology, melanoma examine, pharmacology and inner drugs; and physicians undertaking medical reports in those fields.
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Extra info for Manual of Antisense Methodology
A rapid method for the preparation of a one dimensional sequence-overlapping oligonucleotide library. Nucleot Nucleos 1997;16:835-42 Branch AD. A hitchhiker's guide to antisense and nonantisense biochemical pathways. Hepat%gy 1996;24:1517-29 Branch AD. A good antisense molecule is hard to find. Trends Biochem Sci 1998;23:45-50 Brinkmeier H, Schu B, Seliger H, Kurz LL, Buchholz C, Rudel R. Antisense oligonucleotides discriminating between two muscular Na+ channel isoforms. Biochem Biophys Res Commun 1997;234:235-41 Boiziau C, Kurfurst R, Cazenave C, Roig V, Thuong NT, Toulme 11.
Concentration dependence In a dose response curve a clear dependence of the antisense effect on the oligonucleotide concentration should be visible. only the polarity of the 36 How to choose optimal targets in mRNA Inverted or sense oligonucleotides are not recommended. Similar control reactions can be used to verify the antisense effect in cells or in vivo. 5 ALTERNATIVE METHODS FOR THE SCREENING OF TARGET SITES RNase H assay to screen for accessible target sites on the RNA One major mechanism even if not the exclusive one of antisense inhibition of translation is the degradation of RNA in the mRNAlantisense oligonucleotide duplex by RNase H.
Beaucage SL, Caruthers MH. Deoxynucleoside phosphoramidites-a new class of key intermediates for deoxypolynucleotide synthesis. Tetrahedron Lell 1981 ;22: 1859-62. Beaucage SL, Caruthers MH. "The Chemical Synthesis of DNA/RNA", Bioorganic Chemistry: Nucleic Acids 1996;36-74. SM Hecht, ed. Oxford University Press. Boal JH. Wilk A, Harindranath N, Max EE. Kempe T, Beaucage SL. Cleavage of oligodeoxyribonucleotides from controlled-pore glass supports and their rapid deprotection by gaseous amines.