Antisense Technology, Part A, General Methods, Methods of by John N. Abelson, Melvin I. Simon, M. Ian Phillips

By John N. Abelson, Melvin I. Simon, M. Ian Phillips

Antisense expertise is the facility to govern gene expression inside of mammalian cells offering robust experimental methods for the examine of gene functionality and gene law. for instance, equipment that inhibit gene expression allow stories which probe the conventional functionality of a selected product inside a mobilephone. Such technique can be utilized in lots of disciplines akin to pharmacology, oncology, genetics, cellphone biology, developmental biology, molecular biology, biochemistry, and neurosciences. This quantity can be a really vital instrument in biomedical-oriented learn. The seriously acclaimed laboratory common for greater than 40 years, Methods in Enzymology is among the so much hugely revered courses within the box of biochemistry. because 1955, each one quantity has been eagerly awaited, often consulted, and praised via researchers and reviewers alike. Now with greater than three hundred volumes (all of them nonetheless in print), the sequence comprises a lot fabric nonetheless proper today--truly a necessary booklet for researchers in all fields of lifestyles sciences.

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Griffey, E. A. Lesnik, and B. P. Monia, Biochem. J. 312, 599 (1995). 68 Using a series of noncleavable substrates and Michaelis–Menten analyses, we were able to evaluate both binding and cleavage. We showed that, in fact, E. coli RNase H1 is a double-strand RNA-binding protein. 6 ȐM; the Kd for a DNA duplex was 176 ȐM; and the Kd for single-strand DNA was 942 ȐM. In contrast, the enzyme could only cleave RNA in an RNA–DNA duplex. Any 2Ј modification in the antisense drug at the cleavage site inhibited cleavage, but a significant charge reduction and 2Ј modifications were tolerated at the binding site.

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P. 79. CRC Press, Boca Raton, FL, 1989. 56 C. A. -C. Cheng, Science 261, 1004 (1993). 57 C. Cazenave, C. A. Stein, N. Loreau, N. T. Thuong, L. M. Neckers, C. Subasinghe, C. Hele´ne, J. S. -J. Toulm, Nucleic Acids Res. 17, 4255 (1989). 58 R. S. Quartin, C. L. Brakel, and J. G. Wetmur, Nucleic Acids Res. 17, 7253 (1989). 59 P. J. Furdon, Z. Dominski, and R. Kole, Nucleic Acids Res. 17, 9193 (1989). 60 P. S. Eder and J. A. Walder, J. Biol. Chem. 266, 6472 (1991). 61 B. P. Monia, E. A. Lesnik, C.

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