Pharmaceuticals

GenomONE-Si enables efficient siRNA/miRNA transfection in a variety of difficult-to-transfect cells. Although GenomONE-Neo can also be used for siRNA/miRNA transfection, GenomONE-Si is expected to have a greater knockdown effect with simple operability.

*This product is sold for research purpose only. It may not be used for treatment or other clinical purposes or for intra- and extracorporeal diagnosis in humans or animals.

Features of GenomONE-Si

• Optimal for transfection of synthetic oligo-type siRNA/miRNA
• Simple operability (completion of transfection within 5 to 10 minutes)
• Applicable also to suspended immune cells, transfection into which is usually difficult
  (e.g., Raji, THP-1, U-937, Jurkat, HL-60, Mouse primary T cell)
• Compatible for in vivo delivery
• Optimal for rapid screening of a large number of test samples (HTS：High-throughput screening)

siRNA transfection into mouse primary T cells
Foxp3 induction by knockdown of CDK8/19 expression

Excerpt from "siRNA transfection into mouse primary T cells -Foxp3 induction by knockdown of CDK8/19 expression-"

Fig.1 Knockdown of CDK8 or 19 expression in mouse primary T cells.

Fig.2 Effects of CDK8/19 knockdown on Foxp3 expression in mouse primary T cells.

Transfection of CDK8/19-targeting siRNA using GenomONE-Si suppressed CDK8/19 expression in mouse primary T cells (Fig. 1). Knockdown of CDK8/19 expression facilitated Treg conversion (Fig. 2).

【Data】
Dr. Norihisa Mikami
Department of Experimental Immunology, WPI Immunology Frontier Research Center, Osaka University

【Related article】
Sci. Immunol, 4, eaaw2707 (2019).

Cyclophilin B miRNA transfection in mouse primary T cell and B cell

One day after stimulation of mouse splenocytes with PMA/ionomycin, T cells were isolated. One day after stimulation of mouse splenocytes with anti-CD40/LPS, B cells were isolated. Two days after transfection of cyclophilin B miRNA using GenomONE-Si, mRNA expression of cyclophilin B was measured by qRT-PCR.

CDC2 siRNA transfection in difficult-to-transfect cells

In difficult-to-transfect cells, Raji and HL-60, CDC2 siRNA was transfected using GenomONE-Si, product X, or product R (competitors). Two days after transfection, mRNA expression of CDC2 was measured by qRT-PCR.

Eg5 siRNA transfection in difficult-to-transfect cells

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• Viable cell number（%）=（1 – Eg5 siRNA A450 / Non-targeting siRNA A450）×100

In difficult-to-transfect cells, specifically U-937, Jurkat, and THP-1 cells, Eg5 siRNA was transfected using GenomONE-Si, product X, or product R (competitors). Two days after transfection, knockdown effect(mitotic arrest, apoptosis) was assessed by WST-8 (a cell counting assay, A450).

Successfully transfected cells

A549, C2C12, Caco-2, CMK6G3(Monkey ES cell), COLO 201, D54 MG, Du-145, H1299, HCT 116, HeLa, HeLa S3, Hep G2, HL-60, HMVEC-dLyNeo, HT-29, HuH-6, HuH-7, HUVEC, INS-1E, J774A.1, Jurkat, K-562, MCF-10A, ME-180, MIA PaCa-2, MIN6, mpkCCD(c14), NCI-H929, NIH-3T3, Raji, Ramos, RAW 264.7, Reh, SK-CO-1, SMMC-7721, SW-480, SW-620, TE-13, TF-1a, THP-1, U251 MG, U-937, YN-1, Human primary monocyte, Human primary T cell, Normal human epidermal melanocyte, Mouse primary B cell, Mouse primary T cell, Mouse primary macrophage, Mouse primary granulosa cell, Mouse primary alveolar type 2 cell, Mouse primary mast cell, Rat primary cardiac myocyte, Rat primary granulosa cell

Related technical information

• References

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  Instruction manual

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  siRNA/miRNA transfection in immune cells
  Data sheet

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  High-throughput screening
  Data sheet