Altogen Labs provides development of stable cell line service by transforming your cell line to stably express vector or gene of interest (including RNAi and Tet-inducible cell lines). Cloning plasmid DNA encoding gene of interest, correct promoter, and antibiotic resistance gene is very important part of the process. Plasmid DNA construct should encode at least one antibiotic resistance gene to enable drug selection.
After cloning of plasmid DNA is complete, transfected genetic material can be expressed in the target cells either transiently or permanently depending on the methods utilized and the experimental questions being investigated. Transient transfections are used most commonly to analyze the short term impact of altered gene or protein expression. Plasmid DNA (pDNA) can integrate into host cell line genome and be expressed in a stable manner. Messenger RNA (mRNA) and small RNAs, such as short interfering RNA (siRNA) and microRNA (miRNA), are introduced via transient transfection process and gene products are expressed in the target cells however the nucleic acids do not integrate into the host cell genome. Therefore, gene product expression is transient and typically results in high expression levels that persist for 24-72 hours when RNA is transfected, or 48-96 hours following transient DNA transfection.
Conversely, in order to analyze the long term impact of altered gene or protein expression investigators typically utilize stable transfection protocols to develop stable cell lines. In a subpopulation of transfected cells, whether the desired effect is a stable or transient transfection, the transfected genetic material will integrate into the genome. In order to create stable cell lines, investigators will take advantage of this natural occurrence, and introduce the gene of interest along with a selectable marker. Therefore growth of transfected cells, in the presence of a selecting agent, will enable the subpopulation of cell where the exogenous genetic material has been incorporated into the genome to persist while the remaining cells undergo selection. Utilizing this method, investigators are able to develop cells that permanently express specific genes through their incorporation in the cellular genome. This method is widely used for protein manufacturing in biotechnology and pharma industry.
Altogen Labs provide development of stable cell lines service in just 28 days: Receive instant quote for 28-days stable cell line development
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