Lessons from Nature: microRNA-based shRNA libraries.

Nat Methods 2006 Sep;3(9):707-14

Cold Spring Harbor Laboratory, Watson School of Biological Sciences, Howard Hughes Medical Institute, 1 Bungtown Road, Cold Spring Harbor, New York 11724, USA.

Loss-of-function genetics has proven essential for interrogating the functions of genes and for probing their roles within the complex circuitry of biological pathways. In many systems, technologies allowing the use of such approaches were lacking before the discovery of RNA interference (RNAi). We have constructed first-generation short hairpin RNA (shRNA) libraries modeled after precursor microRNAs (miRNAs) and second-generation libraries modeled after primary miRNA transcripts (the Hannon-Elledge libraries). These libraries were arrayed, sequence-verified, and cover a substantial portion of all known and predicted genes in the human and mouse genomes. Comparison of first- and second-generation libraries indicates that RNAi triggers that enter the RNAi pathway through a more natural route yield more effective silencing. These large-scale resources are functionally versatile, as they can be used in transient and stable studies, and for constitutive or inducible silencing. Library cassettes can be easily shuttled into vectors that contain different promoters and/or that provide different modes of viral delivery.

Download full-text PDF

Source
http://dx.doi.org/10.1038/nmeth923DOI Listing
September 2006

Publication Analysis

Top Keywords

second-generation libraries
8
libraries modeled
8
shrna libraries
8
libraries
6
modeled primary
4
primary mirna
4
stable studies
4
libraries arrayed
4
versatile transient
4
mirna transcripts
4
hannon-elledge libraries
4
transcripts hannon-elledge
4
mirnas second-generation
4
libraries libraries
4
transient stable
4
precursor micrornas
4
constitutive inducible
4
first-generation short
4
inducible silencing
4
constructed first-generation
4

Altmetric Statistics

References

(Supplied by CrossRef)

PD Zamore et al.
Cell 2000

AJBDC Hamilton et al.
Science 1999

SM Hammond et al.
Nature 2000

E Bernstein et al.
Nature 2001

SM Hammond et al.
Science 2001

T Tuschl et al.
Genes Dev. 1999

J Liu et al.
Science 2004

J-J Song et al.
Science 2004

DS Schwarz et al.
Cell 2003

A Khvorova et al.
Cell 2003

RC Lee et al.
Cell 1993

Similar Publications