By John N. Abelson, Melvin I. Simon, M. Ian Phillips
Antisense expertise is the facility to govern gene expression inside mammalian cells delivering robust experimental methods for the examine of gene functionality and gene rules. for instance, tools that inhibit gene expression allow reports which probe the conventional functionality of a particular product inside a cellphone. Such technique can be utilized in lots of disciplines similar to pharmacology, oncology, genetics, mobile biology, developmental biology, molecular biology, biochemistry, and neurosciences. This quantity may be a very very important software in biomedical-oriented learn. The seriously acclaimed laboratory common for greater than 40 years, Methods in Enzymology is among the such a lot hugely revered guides within the box of biochemistry. considering 1955, every one quantity has been eagerly awaited, often consulted, and praised by means of researchers and reviewers alike. Now with greater than three hundred volumes (all of them nonetheless in print), the sequence includes a lot fabric nonetheless appropriate today--truly a vital ebook for researchers in all fields of lifestyles sciences.
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Additional info for Antisense Technology, Part A, General Methods, Methods of Delivery, and RNA Studies
It is likely that ISIS 1939 destabilizes the RNA and activates RNase H. In contrast, ISIS 1570, an 18-mer phosphorothioate that is complementary to the translation initiation codon of the ICAM-1 message, inhibited production of the protein, but caused no degradation of the RNA. 66 Given the emerging role of chimeric oligonucleotides with modiﬁcations in the 3Ј- and 5Ј-wings designed to enhance afﬁnity for the target RNA and nuclease stability and a DNA-type gap to serve as a substrate for RNase H, studies focused on understanding the effects of various modiﬁcations on the efﬁciency of the enzyme(s) are also of considerable importance.
Biophys. Methods 20, 259 (1990). 99 T. Miyao, Y. Takakura, T. Akiyama, F. Yoneda, H. Sezaki, and M. Hashida, Antisense Res Dev. 5, 115 (1995). 22 GENERAL METHODS  strategies in which oligonucleotides are modiﬁed at only the 3Ј and 5Ј terminus as a means of enhancing stability have not proven to be successful. In Vitro Cellular Uptake. 102,103 Uptake is time and temperature dependent. 16 Nor are the factors that result in differences in the uptake of different sequences of oligonucleotide understood.
CRC Press, Boca Raton, FL, 1989. 46 R. J. -L. Dirksen, in ‘‘Nucleases’’ (S. M. Linn and R. J. ), p. 211. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1985. 47 C. Crum, J. D. Johnson, A. Nelson, and D. Roth, Nucleic Acids Res. 16, 4569 (1988). 48 M. T. Haeuptle, R. Frank, and B. Dobberstein, Nucleic Acids Res. 14, 1427 (1986). 49 H. Donis-Keller, Nucleic Acids Res. 7 (1979). 52,53 Additionally, backbone modiﬁcations inﬂuence the ability of oligonucleotides to activate RNase H. 63–65 In fact, direct proof that RNase 50 A.