RNA interference rescue by bacterial artificial chromosome transgenesis in mammalian tissue culture cells

by: Ralf Kittler, Laurence Pelletier, Chunling Ma, Ina Poser, Steffi Fischer, Anthony A Hyman, Frank Buchholz

PNAS, Vol. 102, No. 7. (15 February 2005), pp. 2396-2401.

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Abstract

RNA interference (RNAi) is a widely used method for analysis of gene function in tissue culture cells. However, to date there has been no reliable method for testing the specificity of any particular RNAi experiment. The ideal experiment is to rescue the phenotype by expression of the target gene in a form refractory to RNAi. The transgene should be expressed at physiological levels and with its different splice variants. Here, we demonstrate that expression of murine bacterial artificial chromosomes in human cells provides a reliable method to create RNAi-resistant transgenes. This strategy should be applicable to all eukaryotes and should therefore be a standard technology for confirming the specificity of RNAi. We show that this technique can be extended to allow the creation of tagged transgenes, expressed at physiological levels, for the further study of gene function. 10.1073/pnas.0409861102

@article{citeulike:1243169, abstract = {RNA interference (RNAi) is a widely used method for analysis of gene function in tissue culture cells. However, to date there has been no reliable method for testing the specificity of any particular RNAi experiment. The ideal experiment is to rescue the phenotype by expression of the target gene in a form refractory to RNAi. The transgene should be expressed at physiological levels and with its different splice variants. Here, we demonstrate that expression of murine bacterial artificial chromosomes in human cells provides a reliable method to create RNAi-resistant transgenes. This strategy should be applicable to all eukaryotes and should therefore be a standard technology for confirming the specificity of RNAi. We show that this technique can be extended to allow the creation of tagged transgenes, expressed at physiological levels, for the further study of gene function. 10.1073/pnas.0409861102}, author = {Kittler, Ralf and Pelletier, Laurence and Ma, Chunling and Poser, Ina and Fischer, Steffi and Hyman, Anthony A. and Buchholz, Frank }, citeulike-article-id = {1243169}, doi = {10.1073/pnas.0409861102}, journal = {PNAS}, keywords = {rnai}, month = {February}, number = {7}, pages = {2396--2401}, posted-at = {2007-09-20 03:48:32}, priority = {2}, title = {RNA interference rescue by bacterial artificial chromosome transgenesis in mammalian tissue culture cells}, url = {http://dx.doi.org/10.1073/pnas.0409861102}, volume = {102}, year = {2005} }

RIS record

TY - JOUR ID - citeulike:1243169 L3 - citeulike-article-id:1243169 TI - RNA interference rescue by bacterial artificial chromosome transgenesis in mammalian tissue culture cells JF - PNAS VL - 102 IS - 7 SP - 2396 EP - 2401 N2 - RNA interference (RNAi) is a widely used method for analysis of gene function in tissue culture cells. However, to date there has been no reliable method for testing the specificity of any particular RNAi experiment. The ideal experiment is to rescue the phenotype by expression of the target gene in a form refractory to RNAi. The transgene should be expressed at physiological levels and with its different splice variants. Here, we demonstrate that expression of murine bacterial artificial chromosomes in human cells provides a reliable method to create RNAi-resistant transgenes. This strategy should be applicable to all eukaryotes and should therefore be a standard technology for confirming the specificity of RNAi. We show that this technique can be extended to allow the creation of tagged transgenes, expressed at physiological levels, for the further study of gene function. 10.1073/pnas.0409861102 KW - rnai AU - Kittler, Ralf AU - Pelletier, Laurence AU - Ma, Chunling AU - Poser, Ina AU - Fischer, Steffi AU - Hyman, Anthony AU - Buchholz, Frank PY - 2005/02/15/ UR - http://dx.doi.org/10.1073/pnas.0409861102 ER -

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