The Human Lincode NR lncRNA RefSeq v65 siRNA library targets long noncoding RNA (lncRNA) transcripts with NR accession number designation; both from lncRNA genes and within protein coding genes. The Lincode siRNA designs and modifications reduce off-target effects while maintaining high silencing potency for high-confidence screening results.
- Patented dual-strand modification pattern on all siRNAs to reduce off-targets
- Sense strand is modified to prevent interaction with RISC and favor antisense strand uptake
- Antisense strand seed region is modified to destabilize off-target activity and enhance target specificity
- Available as SMARTpool siRNA reagents or a Set of 4 siRNAs in 96-well or 384-well plates
This library targets 2022 unique human lncRNA genes and 1439 lncRNA transcripts that are associated with protein-coding gene records for a total of 3461 targets.
For a complete list of target genes in this siRNA Library, please contact Technical Support or your local Sales Representative. For pricing on this library or to request a custom library, please submit a Library Quote Request.
|Storage Conditions||-20 C|
|Stability at Recommended Storage Conditions||At least 12 months|
Detection of effective lncRNA knockdown following application of Lincode siRNA
Knockdown of CDKN2B-AS1 in HeLa cells by Lincode SMARTpool and four siRNAs. All siRNAs were used at 25 nM. LncRNA transcripts were detected with Solaris (sold under the Thermo Scientific brand) qPCR lncRNA Expression Assays and normalized to Non-targeting Control siRNA. Viability was assessed by resazurin assay.
lncRNAs have highly variable expression
Expression analysis of nine lncRNAs varies widely across three human cell lines.
It is of vital importance to determine lncRNA expression in your cells prior to knockdown experiments. lncRNA expression can be expected to vary much more than is typically observed from protein-coding genes like Cyclophilin B (PPIB).
50 ng of total RNA was converted into cDNA using the Maxima First Strand cDNA Synthesis Kit (# K1641). PPIB was used as a reference for typical mRNA expression. Standard RT-PCR cycling conditions resulted in initial Cq values ranging from 22 (PPIB in 293T cells) to 36 (BDNF-AS1 in HeLa cells). lncRNA MLK7-AS1 expression was not detectable in any of the cell lines tested. Cell lines tested include HeLa, HEK293T, Human Neonatal Dermal Fibroblast (hNDF).
Lincode modifications enhance function and reduce off-targets
Lincode siRNA effectively knocks down BDNF-AS1 lncRNA, but not the protein-coding transcript BDNF that is antisense to the lncRNA target. This indicates strand specificity of Lincode siRNAs and effectiveness of ON TARGETplus modifications. siRNAs were applied at the indicated concentrations to hNDF cells. BDNF-AS1 and BDNF transcripts were detected with Solaris (sold under the Thermo Scientific brand) qPCR Expression Assays and normalized to Non-targeting control siRNA. Viability was assessed by resazurin assay and normalized to Untreated.
Sense strand activity is prevented by siRNA modifications
Lincode siRNAs are modified with a proprietary dual-strand modification that improves siRNA functionality. Additionally, off-targets are reduced due to:
- Inactivation of passenger strand activity; driving preferential loading of the guide strand into RISC
- Novel seed region modifications for disruption of microRNA-like off-targets
Lincode siRNAs effectively knockdown target lncRNAs
Knockdown of BDNF-AS1 in hNDF cells by Lincode SMARTpool and four siRNAs. All siRNAs were used at 25 nM. LncRNA transcripts were detected with Solaris (sold under the Thermo Scientific brand) qPCR lncRNA Expression Assays and normalized to Non-targeting Control siRNA. Viability was assessed by resazurin assay.