stealth siRNA GIRK1/KCNJ3

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	The human estrogen non-responsive MDA-MB-453 breast cancer cell line was purchased from the American Type Culture Collection (Rockville, MD, U.S.A.) and was maintained in RPMI 1640 medium supplemented with fetal bovine serum (10%, v/v), L-glutamine (2 mM), 100 U/ml penicillin, and 100 μg/ml streptomycin at 37 °C in an environment of 5% CO2.Three stealth siRNA constructs (Invitrogen, Carlsbad, CA, U.S.A.) were made starting at bases 1104, 1315, and 1490 of the GIRK1 sequence (GenBank # NM_002239). These constructs are: 1104 sense—gga aac aac ugg gau gac uug uca a; 1104 antisense—uug aca agu cau ccc agu ugu uuc c (GC content 44%); 1315 sense—cca gcc aua acu aac agc aaa gaa a; 1315 antisense—uuu cuu ugc ugu uag uua ugg cug g (GC content 40%); 1490 sense—acu ugc cca uga aac uuc aac gaa u; 1490 antisense—auu cgu uga agu uuc aug ggc aag u (GC content 40%). In addition, the low GC RNAi negative control (35%–45% GC) was used as recommended by the manufacturer (Invitrogen). These constructs (200 pMol) were transfected into the MDA-MB-453 breast cancer cell lines using lipofectamine 2000 (Invitrogen).

Protocol tips

The human estrogen non-responsive MDA-MB-453 breast cancer cell line was purchased from the American Type Culture Collection (Rockville, MD, U.S.A.) and was maintained in RPMI 1640 medium supplemented with fetal bovine serum (10%, v/v), L-glutamine (2 mM), 100 U/ml penicillin, and 100 μg/ml streptomycin at 37 °C in an environment of 5% CO2.Three stealth siRNA constructs (Invitrogen, Carlsbad, CA, U.S.A.) were made starting at bases 1104, 1315, and 1490 of the GIRK1 sequence (GenBank # NM_002239). These constructs are: 1104 sense—gga aac aac ugg gau gac uug uca a; 1104 antisense—uug aca agu cau ccc agu ugu uuc c (GC content 44%); 1315 sense—cca gcc aua acu aac agc aaa gaa a; 1315 antisense—uuu cuu ugc ugu uag uua ugg cug g (GC content 40%); 1490 sense—acu ugc cca uga aac uuc aac gaa u; 1490 antisense—auu cgu uga agu uuc aug ggc aag u (GC content 40%). In addition, the low GC RNAi negative control (35%–45% GC) was used as recommended by the manufacturer (Invitrogen). These constructs (200 pMol) were transfected into the MDA-MB-453 breast cancer cell lines using lipofectamine 2000 (Invitrogen).

Publication protocol

The human estrogen non-responsive MDA-MB-453 breast cancer cell line was purchased from the American Type Culture Collection (Rockville, MD, U.S.A.) and was maintained in RPMI 1640 medium supplemented with fetal bovine serum (10%, v/v), L-glutamine (2 mM), 100 U/ml penicillin, and 100 μg/ml streptomycin at 37 °C in an environment of 5% CO2.Three stealth siRNA constructs (Invitrogen, Carlsbad, CA, U.S.A.) were made starting at bases 1104, 1315, and 1490 of the GIRK1 sequence (GenBank # NM_002239). These constructs are: 1104 sense—gga aac aac ugg gau gac uug uca a; 1104 antisense—uug aca agu cau ccc agu ugu uuc c (GC content 44%); 1315 sense—cca gcc aua acu aac agc aaa gaa a; 1315 antisense—uuu cuu ugc ugu uag uua ugg cug g (GC content 40%); 1490 sense—acu ugc cca uga aac uuc aac gaa u; 1490 antisense—auu cgu uga agu uuc aug ggc aag u (GC content 40%). In addition, the low GC RNAi negative control (35%–45% GC) was used as recommended by the manufacturer (Invitrogen). These constructs (200 pMol) were transfected into the MDA-MB-453 breast cancer cell lines using lipofectamine 2000 (Invitrogen).

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing siRNA / miRNA gene silencing Human - MDA-MB-231 GIRK1/KCNJ3 using stealth siRNA GIRK1/KCNJ3 from Thermo Fisher Scientific.

Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for stealth siRNA GIRK1/KCNJ3 below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing siRNA / miRNA gene silencing Human - MDA-MB-231 GIRK1/KCNJ3 using stealth siRNA GIRK1/KCNJ3 from Thermo Fisher Scientific. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.