Stable Expression Cell Lines

Stable Expression Cell Lines

Postby biologist on Fri Oct 02, 2009 10:29 pm

Stably expressing cell lines are useful research models used in basic research, as well as development, creation and production of diagnostic proteins, monoclonal antibodies, and recombinant therapeutic proteins. Because cell lines may also be used in the fields of physiology and toxicology, as well as drug screening, producing stable cell lines is essential to these testing environments.

It takes a very long time to generate a stable cell line, so utilizing techniques and methods to do so save time, money, and effort on the part of researchers and developers when it comes to understanding how cells work, express themselves, and may be utilized in a multitude of studies and protocols.

In the past, the creation of a stably expressed cell line could take up to 30 or 40 weeks, but research and development has enabled stably integrated genes for protein synthesis in state-of-the-art facility procedures that can be realized within a relatively short 4 to 6 week turnaround.
biologist
 
Posts: 44
Joined: Sat Aug 01, 2009 9:48 pm

Re: Stable Expression Cell Lines

Postby biologist on Fri Oct 02, 2009 10:31 pm

Stable cell line creation is utilized for RNAi knockdown as well as gene expression and differentiation studies, inducible expression studies, as well as methods and techniques to utilize the transference of cell lines in vivo. Stable cell lines can be created through transfection of DNA plasmids or shRNA-expressing constructs. Selection or the use of certain virus particles will allow DNA integration into recipient cell genomes and help to reduce the formally long period of time that was necessary for the creation of a stable cell line.

Cell lines that have been stably expressed for the purposes of protein or RNAi sequencing is a precise an exact science that utilizes a variety of approaches. The approach is dependent upon the type of expression (constituted or inducible) that is desired. Results demand large-scale biochemical analysis of fusion proteins as well as monitoring of cell cycle localization.

Stably expressing cell lines are able to encode at least one antibiotic gene that is resistant to various drugs in order to enable drug selection processes. Through this process, transfected genetic materials may be expressed in specific target cells depending on investigative as well as methods utilized to determine short-term impact of protein expression or altered gene expression.
biologist
 
Posts: 44
Joined: Sat Aug 01, 2009 9:48 pm

Re: Stable Expression Cell Lines

Postby postdoc on Sun Nov 27, 2011 9:35 am

Altogen Labs provides Generation of Stable RNAi Cell Lines (stable shRNA Gene Knockdown) - http://altogenlabs.com/services/generation_of_stable_cell_lines/stable_rnai_cell_line_gene_knockdown

biologist wrote:Stable cell line creation is utilized for RNAi knockdown as well as gene expression and differentiation studies, inducible expression studies, as well as methods and techniques to utilize the transference of cell lines in vivo. Stable cell lines can be created through transfection of DNA plasmids or shRNA-expressing constructs. Selection or the use of certain virus particles will allow DNA integration into recipient cell genomes and help to reduce the formally long period of time that was necessary for the creation of a stable cell line.

Cell lines that have been stably expressed for the purposes of protein or RNAi sequencing is a precise an exact science that utilizes a variety of approaches. The approach is dependent upon the type of expression (constituted or inducible) that is desired. Results demand large-scale biochemical analysis of fusion proteins as well as monitoring of cell cycle localization.

Stably expressing cell lines are able to encode at least one antibiotic gene that is resistant to various drugs in order to enable drug selection processes. Through this process, transfected genetic materials may be expressed in specific target cells depending on investigative as well as methods utilized to determine short-term impact of protein expression or altered gene expression.
postdoc
 
Posts: 44
Joined: Mon Oct 26, 2009 8:45 am

Re: Stable Expression Cell Lines

Postby ATGC2001 on Sun Nov 27, 2011 2:46 pm

ATGC2001
 
Posts: 26
Joined: Thu Oct 01, 2009 7:39 am

Re: Stable Expression Cell Lines

Postby postdoc on Tue Oct 30, 2012 5:38 pm

Altogen Launches Custom Biotechnology Research Services: Generation of Stable Cell Lines and RNAi Cell Transfection Services

Altogen Labs announced launch of new contract research services - Generation of Stable Cell Lines in 28 Days. Altogen custom services focus on providing specialized biotechnology and pharmaceutical services, including RNA Interference (RNAi) services, generation of stably-expressing cell lines, assay development, screening and transfection services. In particular, the company will draw upon their unique experience with over hundred cancer cell lines and expertise in polymer and nanoparticle-based gene delivery technologies.

Generation of stably-expressing cancer cell lines and primary cells can be very expensive and time-consuming. Utilizing this method, investigators are able to develop cells that permanently express specific genes through their incorporation in the cellular genome. Altogen custom services offers generation of stable cell lines by transforming the cell line of choice to stably express vector or gene of interest. Optional cloning and shRNA expression services are available. The service turnaround time is typically 3-4 weeks.

Altogen laboratory research services include:

-- Transfection services (transient and stable)
-- Assay development and library screening
-- Cloning and plasmid vector construction
-- Generation of stable cell lines
-- RNAi experiments (siRNA design, synthesis, validation)
-- Gene targeting (mRNA knockdown, shRNA overexpression)
-- Custom services



Altogen is a life sciences company dedicated to the development, marketing and manufacture of In Vivo and In Vitro transfection reagents for intracellular and tissue-targeted delivery of biomolecules. Altogen custom services is a laboratory contract research organization. It provides specialized contract research to pharmaceutical, biotechnology, and other life science organizations worldwide.
postdoc
 
Posts: 44
Joined: Mon Oct 26, 2009 8:45 am

Re: Stable Expression Cell Lines

Postby labmember on Thu Jul 03, 2014 11:00 am

Another application of stable mammalian cell lines is in the production of small hairpin RNA (shRNA: http://en.wikipedia.org/wiki/ShRNA) vectors. These viral vectors produce shRNA, which activates the RNA interference (RNAi) pathway, in a process also called gene silencing or gene knockdown. This phenomenon acts to silence the expression of a specific gene by degrading the corresponding messenger RNA (mRNA). It is triggered when double stranded RNA (such as shRNA), combined with the RNA-induced silencing complex (RISC http://en.wikipedia.org/wiki/RNA-induced_silencing_complex), reduces the protein production of a specific gene or genes. RNAi can be useful in many areas, including analysis of gene function, discovery of gene targets, development of assays, and pharmaceutical compound screening. These cell lines are complex to generate, due to the large number of details involved, including controlling for off-target events, modifying efficiency of silencing, and evaluating promoter compatibility.
labmember
 
Posts: 5
Joined: Tue Jun 24, 2014 2:30 pm


Return to Stable Transfection

Who is online

Users browsing this forum: No registered users and 1 guest

cron