Gene expression/KO- (Transfection, Infection, SiRNA)

Genes can be introduced into cells by suitable molecular biology methods such as electroporation, cationic lipid reagents, or calcium phosphate. A gene can be transfected into cells simply with its regulatory elements or after making a construct to achieve high overexpression levels. Cells can also be transfected with conditionally expressed genes of interest. Transfections in mammalian cells can be transient or permanent. Transient expression lasts only a few days. Permanent expression requires cotransfection with a dominant selectable marker and several rounds of selection for the cell populations that stably integrate the transfected DNA into a cellular gene. This takes approximately three to four weeks.

Functional genomics is a great tool to study dynamics aspects of gene transcription, translation and regulation of gene expression. At MuriGenics we can investigate function of your target at the levels of DNA/genes, RNA transcripts, and protein products.

RNA Isolation and quantification

RNA expression analysis is the first step to look for changes in the cells/tissues upon treatment. At murigenics we can isolate RNA from various tissues including tumors, lungs, skin, etc.. Isolated RNA can be further used for gene expression analysis like qPCR, NanoString or RNAseq.

Gene expression analysis

At RNA level, we can investigate the expression level of various genes. These can be done by routine quantitative PCR or with the use of pathway driven microarrays. We can also process samples for RNA sequencing analysis, which has becoming increasing afforbale with the advancement of sequencing technologies.

At MuriGenics, we can introduce gene of interest into cells by suitable molecular biology methods such as electroporation, cationic lipid reagents, or calcium phosphate. A gene can be transfected into cells simply with its regulatory elements or after making a construct to achieve high overexpression levels. Also, cells can also be transfected with conditionally expressed genes of interest. We can perform transfection experiments in mammalian cells for transient or permanent gene expression. Transient expression lasts only a few days whereas permanent expression requires co-transfection with a dominant selectable marker and several rounds of selection for the cell populations that stably integrate the transfected DNA into a cellular gene.

RNA isolation , reverse transcriptase, RT-PCR,

• RNA interference – Si RNA/shRNA expression knockdowns
• Genome editing of individual and multiples genes – including CRISPR_CAS9
• Episomal detection of miRNA
• RNA expression for pathway analysis – Arrays