RNAi Transfection Optimization Services

BOC Sciences' RNAi transfection optimization services include siRNA delivery optimization and plasmid transfection optimization. As both siRNA and RNA interference vectors need to be transferred into cells in order to function. The efficient transfer of RNA interference products into cells is therefore a critical step for successful RNA interference experiments. Professional siRNA and plasmid transfection techniques can be used to optimize transfection efficiency and enhance the effectiveness of RNA interference.

Introduction

The phenomenon of RNA interference (RNAi), which is frequently present in living organisms, is a sequence-specific post-transcriptional silencing process of genes that is sparked by double-stranded RNA homologous to the sequence of the target gene. The cytoplasmic enzyme Dicer breaks down dsRNA into siRNAs, which include 21-25 nucleotides. The RNA-induced silencing complex (RISC), which specifically binds to the homologous area of exogenous gene expression mRNAs and cleaves the mRNAs at the binding site, is created in vivo when the siRNAs attach to proteins. The degraded, damaged mRNAs are then used to prevent the associated gene's post-transcriptional mechanism for silencing from working.

RNAi has emerged as a crucial tool for understanding gene function and will be crucial in the treatment of viral diseases, genetic disorders, and oncological diseases by specifically inhibiting the overexpression of genes like HIV, hepatitis, oncogenes, and other related genes and keeping them in a silent or dormant state.

The application of RNAi technology is limited by transfection. The most common and crucial transfection for RNAi technology is siRNA transfection. siRNA transfection experiments involve the following important parameters:

• Health of cultured cells
• Transfection methods
• Transfection conditions
• Quality and quantity of siRNA

Service Content

Service Process

• Culture the cells
• Optimize transfection conditions: observe the transfection efficiency under fluorescence microscopy or assess transfection efficiency with flow cytometry
• Design a set of RNA interference sequences based on the target gene sequences
• Evaluate RNA interference efficiency by fluorescence PCR or Western blot
• (Optional) Screen cell lines for stable expression of shRNA or miRNA using appropriate antibiotics

Customer Provided

• Cell line information and detailed cell culture procedures
• Target gene GeneBank number
• Antibodies (if Western blot assay is required)

Experiments provided

• Optimization of transfection efficiency analysis
• Analysis of RNA interference efficiency

Quality Assurance

• 3 siRNAs designed for one target gene can guarantee that 1 of them interferes with greater than 70% efficiency (under the premise that transfection efficiency is greater than 80%).
• 3 miRNA interference vectors designed and constructed for one target gene can guarantee that 1 of them interferes with greater than 70% efficiency (under the premise that the transfection efficiency is greater than 80%)