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Transient RNA interference in hematopoietic progenitors with functional consequences
Author(s) -
Oliveira Daniela M.,
Goodell Margaret A.
Publication year - 2003
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/gene.10212
Subject(s) - gene knockdown , small interfering rna , rna interference , haematopoiesis , biology , transfection , progenitor cell , electroporation , microbiology and biotechnology , rna , gene expression , gene silencing , small hairpin rna , stem cell , gene , genetics
Summary: Short interfering (si) RNAs have now been shown to inhibit gene expression in several species, including mammals (Elbashir et al. : Nature 411:494–498, 2001; Fire et al. : Nature 391:806–811, 1998). RNA inhibition in primary cells such as stem cells would facilitate rapid gene discovery in a postgenome era. While retroviruses can deliver siRNA expression cassettes for stable expression (Barton and Medzhitov: Proc Natl Acad Sci USA 99:14943–14945, 2002; Paddison et al. : Proc Natl Acad Sci USA 99:1443–1448, 2002; Rubinson et al. : Nat Genet 33:401–406, 2003), an efficient method for direct transfer of siRNA to stem cells is still lacking. Here, we established electroporation to deliver siRNA to hematopoietic progenitors. On average, at least 80% of cells take up the RNA, and these display nearly 100% knockout of marker gene expression at both the RNA and protein level. Moreover, knockdown of the hematopoietic regulator, CD45, results in 3‐fold more hematopoietic colonies in a progenitor assay. These results demonstrate that transient transfection of siRNA to primary cells can have substantial functional consequences. This technology may be applicable to a variety of primary cell types. genesis 36:203–208, 2003. © 2003 Wiley‐Liss, Inc.