Human sigenome smartpool library

Manufactured by Horizon Discovery

The Human siGenome SMARTpool library is a collection of small interfering RNA (siRNA) reagents designed for gene knockdown studies in human cells. The library targets thousands of human genes and is intended for use in high-throughput screening and gene function analysis applications.

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Lab products found in correlation

Opti mem medium, by Thermo Fisher Scientific (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Mithras lb941, by Berthold Technologies (1 mentions)

Close spelling variants of this product

SMARTpool (292 citations) SiGENOME SMARTpool (203 citations) SiGENOME Human SMARTpool (8 citations) SiGENOME library (4 citations) SiGENOME SMARTpools library (2 citations)

2 protocols using human sigenome smartpool library

Generating Stable Gene Knockdown and Overexpression Cell Lines

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siRNAs targeting MEK1 and MEK2 from Human siGenome SMARTpool library (Dharmacon) were used in siRNA-mediated gene silencing. H358 cells were transfected using DharmaFECT transfection reagent #1 and 25 nM siRNA. The lentiviral-based RNAi Consortium (TRC) human genome-wide shRNA collection (TRCHs1.0) was used in making gene knockdown cell lines. Individual lentiviral plasmids containing shRNAs against JUN or DUSP4 were collected from TRC library. The CCSB-Broad lentiviral collection of human ORFs was used in making gene overexpression cell lines. The lentiviruses were produced as described at http://www.broadinstitute.org/rnai/public/resources/protocols. In brief, HEK293T cells were transfected with lentiviral vectors using calcium phosphate method. Lentiviral supernatants were collected and transduced into target cells with polybrene (1 mg/mL). Stable gene knockdown or overexpression cell lines were selected with puromycin (2 μg/mL) or Blasticidin (10 μg/mL).

Xue Z., Vis D.J., Bruna A., Sustic T., van Wageningen S., Batra A.S., Rueda O.M., Bosdriesz E., Caldas C., Wessels L.F, & Bernards R. (2018). MAP3K1 and MAP2K4 mutations are associated with sensitivity to MEK inhibitors in multiple cancer models. Cell Research, 28(7), 719-729.

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Systematic Screening of Inflammation Genes

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The human siGENOME SMART pool library targeting 1027 inflammation-related genes were obtained from Dharmacon. For the siRNA screens, transfection mixtures were 50nM siRNA, 100ng OCT4 reporter constructs (QIAGEN) and 0.3 ul Lipofectamine 2000 (Invitrogen) in 50 ul opti-MEM medium (Invitrogen), three replicates were used for each siRNA. In each plate, we used both POU5FI (5460) siRNA-SMART pool Dharmacon M-019591-03-0005 and SiGENOME Human PRDM14 (63978) siRNA-SMART pool as positive controls [21 (link)]. HMLE-snail cell were plated in 50ul opti-MEM medium with a density of 10000 cells per well in 96-well plate. Dual Luciferase reporter assay was carried out for medium-throughput readout (Promega), and the level of OCT4 reporter gene expression was measured using a microplate reader (Mithras LB941, Berthold Technologies). OCT4 reporter values were normalized to the expression of the Renilla luciferase reporter. Fold change value of each siRNA sample was calculated. The distribution of the datasets was determined to be closed to normal distribution, so the Student pair-t test between each siRNA group and the relative negative control group was performed. The cut-off values for the screen are Fold change <0.5 or >2.0, and P<0.05.

Shen W., Xie J., Zhao S., Du R., Luo X., He H., Jiang S., Hao N., Chen C., Guo C., Liu Y., Chen Y., Sun P., Yang S., Luo N., Xiang R, & Luo Y. (2018). ICAM3 mediates inflammatory signaling to promote cancer cell stemness. Cancer letters, 422, 29-43.

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