Antisense Therapeutics 2nd edition (Methods in Molecular by M. Ian Phillips (Editor)

By M. Ian Phillips (Editor)

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By M. Ian Phillips (Editor)

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Chem. 254, 10,180– 10,183. 50. 50 Elbashir, S. , et al. (2001) Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 411, 494–498. 51. 51 McManus, M. T. and Sharp, P. A. (2002) Gene silencing in mammals by small interfering RNAs. Nat. Rev. 3, 737–747. 52. 52 Shi, Y. (2003) Mammalian RNAi for the masses. Trends Genet. 19, 9–12. 53. , Lee, S. , et al. (2003) RNA interference targeting Fas protects mice from fulminant hepatitis. Nat. Med. 9, 347–351. 54. 54 Sorensen, D.

Ribozymes recognize specific RNA sequences and then catalyze a site-specific phosphodiester bond cleavage within the target molecule. Following cleavage, the ribozyme releases itself and binds to another target molecule, repeating the process. The cellular consequence varies depending on the setting. There are many naturally occurring ribozymes, including in plant viroids, ribosomes, self-splicing introns, and the RNA portion of RNase P. In plant and animal cells, as well as in viruses, ribozymes are necessary for some normal cellular processes such as transcription.

The ribozyme structure can be engineered to create an intermolecular cleaving ribozyme consisting of two singlestranded antisense arms surrounding the catalytic core and helix 2 so that it will cleave within a different RNA molecule. Because RNA often folds into 20 Zhang et al. Fig. 3. Schematic of a natural hairpin ribozyme. Hairpin ribozymes consist of 4 helices, formed by complementary base pairing, which are connected by single stranded regions. The small loop at the base of the ribozyme is removed to generate a trans-cleaving hairpin ribozyme; helices 1–4 then form portions of the antisense arms.

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