Until recently, the identification of synthetic lethal interactions in human cells was based on screens using chemical libraries or on inferred synthetic lethality in model organisms such as S. cerevisiae[ 1].
The catalytic rates of hydrolysis for substrates with unnatural amino acids designed based on library screening were significantly improved, proving the utility of this approach.
After identification of the genes of interest based on the library screening, a series of RT-qPCR was performed that aimed at investigating the transcription levels of the genes both in vitro cultures and in the bacteria in the organs.
A recent study based on BAC library screening and proteomics analysis showed that Glu-A3, Glu-B3, and Glu-D3 in the Chinese bread wheat cultivar Xiaoyan 54 contain 4, 3, and 7 genes, respectively [ 16].
Many C. elegans genes have been associated with phenotypes due to the results of reverse genetic screens based on RNAi libraries.
It is therefore usually recommended to use some small compositional nanomaterial libraries to perform initial toxicity screening, based on which combinatorial libraries are then introduced for more in-depth studies.
Another strategy, with a proven success in the development of inhibitors of for example HIV, herpes and HCV, has been based on the screening of large libraries of molecules.
For example, based on the screening of genomic libraries with a conserved retrotransposon probe, Hirochika and his colleagues [ 24] made an estimate of ~1000 retrotransposons totally in the rice genome, which fall into 32 families.
An antibody screening based on a library of 45 antibodies directed against surface epitopes including published stem cell and/or cancer stem cell markers (Additional file 2: Table S1) was performed to identify novel biomarkers for breast cancer cell subpopulations resistant to chemotherapeutic treatment.
In this study, we have developed a protocol to identify novel regulators of mammary stem/progenitor cells using freshly isolated MaSC-enriched cells for a functional RNAi screen based on pooled shRNA libraries.
