RNAi in Human Cells: Basic Structural and Functional Features of Small Interfering RNA

by: Ya-Lin Chiu, Tariq M Rana

Molecular Cell, Vol. 10, No. 3. (September 2002), pp. 549-561.

View FullText article

DOI, ScienceDirect

Reviews [Write a review of this article]

There are no reviews of this article

Find related articles from these CiteULike users

tardigrade

Find related articles with these CiteULike tags

no-tag

Abstract

We investigated the mechanism of RNA interference (RNAi) in human cells. Here we demonstrate that the status of the 5′ hydroxyl terminus of the antisense strand of a siRNA determines RNAi activity, while a 3′ terminus block is tolerated in vivo. 5′ hydroxyl termini of antisense strands isolated from human cells were phosphorylated, and 3′ end biotin groups were not efficiently removed. We found no requirement for a perfect A-form helix in siRNA for interference effects, but an A-form structure was required for antisense-target RNA duplexes. Strikingly, crosslinking of the siRNA duplex by psoralen did not completely block RNA interference, indicating that complete unwinding of the siRNA helix is not necessary for RNAi activity in vivo. These results suggest that RNA amplification by RNA-dependent RNA polymerase is not essential for RNAi in human cells.

@article{citeulike:402347, abstract = {We investigated the mechanism of RNA interference (RNAi) in human cells. Here we demonstrate that the status of the 5\′ hydroxyl terminus of the antisense strand of a siRNA determines RNAi activity, while a 3\′ terminus block is tolerated in vivo. 5\′ hydroxyl termini of antisense strands isolated from human cells were phosphorylated, and 3\′ end biotin groups were not efficiently removed. We found no requirement for a perfect A-form helix in siRNA for interference effects, but an A-form structure was required for antisense-target RNA duplexes. Strikingly, crosslinking of the siRNA duplex by psoralen did not completely block RNA interference, indicating that complete unwinding of the siRNA helix is not necessary for RNAi activity in vivo. These results suggest that RNA amplification by RNA-dependent RNA polymerase is not essential for RNAi in human cells.}, author = {Chiu, Ya-Lin and Rana, Tariq M. }, citeulike-article-id = {402347}, doi = {http://dx.doi.org/10.1016/S1097-2765(02)00652-4}, journal = {Molecular Cell}, month = {September}, number = {3}, pages = {549--561}, posted-at = {2005-11-21 05:16:27}, priority = {2}, title = {RNAi in Human Cells: Basic Structural and Functional Features of Small Interfering RNA}, url = {http://dx.doi.org/10.1016/S1097-2765(02)00652-4}, volume = {10}, year = {2002} }

RIS record

TY - JOUR ID - citeulike:402347 L3 - citeulike-article-id:402347 TI - RNAi in Human Cells: Basic Structural and Functional Features of Small Interfering RNA JF - Molecular Cell VL - 10 IS - 3 SP - 549 EP - 561 N2 - We investigated the mechanism of RNA interference (RNAi) in human cells. Here we demonstrate that the status of the 5′ hydroxyl terminus of the antisense strand of a siRNA determines RNAi activity, while a 3′ terminus block is tolerated in vivo. 5′ hydroxyl termini of antisense strands isolated from human cells were phosphorylated, and 3′ end biotin groups were not efficiently removed. We found no requirement for a perfect A-form helix in siRNA for interference effects, but an A-form structure was required for antisense-target RNA duplexes. Strikingly, crosslinking of the siRNA duplex by psoralen did not completely block RNA interference, indicating that complete unwinding of the siRNA helix is not necessary for RNAi activity in vivo. These results suggest that RNA amplification by RNA-dependent RNA polymerase is not essential for RNAi in human cells. AU - Chiu, Ya-Lin AU - Rana, Tariq PY - 2002/09// UR - http://dx.doi.org/10.1016/S1097-2765(02)00652-4 ER -

RIS BibTeX