Targeted gene knockout in mammalian cells by using engineered zinc-finger nucleases

davidd submitted, created time 8 months 2 weeks (www.pnas.org)

Gene knockout is the most powerful tool for determining gene function or permanently modifying the phenotypic characteristics of a cell. Existing methods for gene disruption are limited by their efficiency, time to completion, and/or the potential for confounding off-target effects. Here, they demonstrate a rapid single-step approach to targeted gene knockout in mammalian cells, using engineered zinc-finger nucleases (ZFNs).

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Knockdown of TNFR1 by the sense strand of an ICAM-1 siRNA: dissection of an off-target effect

davidd submitted, created time 10 months 6 days (nar.oxfordjournals.org)

This may be the first example in which the off-target effect of an siRNA is actually responsible for the anticipated effect by acting to reduce expression of a protein (TNFR1) that normally regulates expression of the intended target (ICAM-1).

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Reversible gene knockdown in mice using a tight, inducible shRNA expression system

channel submitted, created time 1 year 9 months (nar.oxfordjournals.org)

RNA interference through expression of short hairpin (sh)RNAs provides an efficient approach for gene function analysis in mouse genetics. Techniques allowing to control time and degree of gene silencing in vivo, however, are still lacking. Here we provide a generally applicable system for the temporal control of ubiquitous shRNA expression in mice. Depending on the dose of the inductor doxycycline, the knockdown efficiency reaches up to 90%

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