Do you have any questions?
Find the ideal kit for your application:
SLAMseq Anabolic Kinetic RNA-Seq
SLAMseq Anabolic RNA-Seq labels newly synthesized (nascent) transcripts in cell culture experiments and detects an increasing signal of nascent RNA as they are being synthesized.
Accurate Time-resolved Analysis of RNA Synthesis
Tracking nascent RNA over time allows to determine RNA synthesis rates and delivers accurate time-resolved information of RNA synthesis in response to a stimulus. SLAMseq allows to demonstrate treatment effects with higher resolution compared to conventional methods. SLAMseq Anabolic RNA-Seq is applicable for research purposes, such as gene regulation studies or deciphering cell signaling pathways, or during drug discovery and development processes, such as target identification and validation or biomarker discovery.
Performance
SLAMseq delivers precise measurements for global transcript biosynthesis rates in living cells and allows to distinguish primary and secondary effects of treatments.
Anabolic Kinetics Labeling of RNA
The SLAMseq Anabolic Kinetics Kit differentiates between nascent and existing RNA. At t0, modified nucleotides (S4U) are added to cell culture media, which results in labeling of newly synthesized RNA. Existing RNA remains unlabeled. At tx, the RNA synthesis is stopped by cell lysis and RNA isolation. Sampling at different intervals, tx, allows for measurement of transcript synthesis rates (Fig. 1). Specific measurement of nascent RNA levels provides insights into transcriptome-wide RNA synthesis dynamics.
Figure 1 | Anabolic kinetics labeling experiment time course. Culturing cells with S4U-containing media (extracellular [S4U], solid green line) changes the intracellular S4U concentration (dashed green line). Nascent RNA will be labeled starting from t0. Time course measurements determine RNA synthesis rates. Transcripts with fast (black solid line) and slow (gray solid line) synthesis rates can be distinguished by relative differences in the increase in S4U detection over time. S4U levels for individual transcripts are measured by counting sequencing reads with T > C conversions.
Assess Nascent RNA Levels and RNA Synthesis Rates
Distinguish newly synthesized (nascent) RNA from total RNA using SLAMseq Anabolic Kinetic 3’ mRNA-Seq with QuantSeq FWD V2 for cost-efficient RNA-Seq analysis of RNA synthesis, or the SLAMseq Kinetics Kit – Anabolic Kinetics Module paired with other library prep kits. Profile nascent RNA levels at specified time points for differential expression analyses and perform time course S4U labeling experiments to map RNA synthesis dynamics for individual transcripts.
Analyze Transcriptome-Wide Expression Dynamics
SLAMseq resolves transcript expression dynamics on a transcriptome-wide scale. Individual transcript RNA synthesis rates can be measured directly (Fig. 3). RNA synthesis rates are depending on gene regulation processes, such as blockers, enhancer, cofactors, and are transcript specific.
Figure 3 | S4U labeling kinetics experiments reveal individual RNA synthesis rates of four gene loci.
Identify Direct Transcriptional Targets of Any Gene
Combining SLAMseq with protein modulators, or drug treatments distinguishes direct (primary) and indirect (secondary) target responses (Fig. 4). Use SLAMseq to dissect signaling pathways underlying biological processes and characterize drug-target responses on the transcriptional level (Muhar, M et al., 2018).
Figure 4 | S4U labeling followed by drug treatment and early-stage sampling (time points: t1 – t2) measures mRNA stability changes to map sequential drug responses.
Find more information about SLAMseq data analysis here or contact us at support@lexogen.com.
Workflow
SLAMseq Anabolic Kinetic RNA-Seq allows to detect RNA synthesis rates over time in cell culture experiments in response to a stimulus, e.g., a fast-acting compound or drug candidate. Optimal concentration of labeling reagent must be determined during exploring phase to limit cytotoxicity.
FAQ
Frequently Asked Questions
Access our frequently asked question (FAQ) resources via the buttons below.
Please also check our General Guidelines and FAQ resources!
How do you like the new online FAQ resource? Please share your feedback with us!
Downloads
Safety Data Sheet
If you need more information about our products, please contact us through support@lexogen.com or directly under +43 1 345 1212-41.
Ordering Information
First time user of SLAMseq?
First Time User? We’re excited to offer you an exclusive introductory offer.
Not sure how to get started with your SLAMseq experiment?
Try the SLAMseq product configurator and find the ideal setup for your application!
Buy from our Webstore
Need a web quote?
You can generate a web quote by Register or Login to your account. In the account settings please fill in your billing and shipping address. Add products to your cart, view cart and click the “Generate Quote” button. A quote in PDF format will be generated and ready to download. You can use this PDF document to place an order by sending it directly to sales@lexogen.com.
Web quoting is not available for countries served by our distributors. Please contact your local distributor for a quote.
