Journal of Glycobiology
Open Access

Analysis of Glycosylation Mutants in Cultured Mammalian Cells.

Burygin Sergei^{* *}Correspondence: Burygin Sergei, Biofunctional Chemistry Laboratory, A. Butlerov Institute of Chemistry, Kazan Federal University, Kazan, Russia, Email:

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Description

Cultured mammalian cells refer to animal cells that have been removed from an organism and are grown and maintained in a laboratory setting using cell culture techniques. These cells are widely used in various fields of biological research, biotechnology, drug development, and medical studies due to their relevance to human biology.

They are often derived from established cell lines, which are populations of cells with specific characteristics that can be cultured and maintained over time. These cell lines can be immortalized, meaning they can divide indefinitely under laboratory conditions. In addition to cell lines, primary mammalian cells can be cultured. These cells are obtained directly from living tissues and have a limited lifespan in culture. They are used for studying primary physiological processes and specific cell types.

Glycosylation mutants

Glycosylation mutants of cultured mammalian cells are cell lines that have been genetically engineered to disrupt or alter specific glycosylation pathways. These mutants are valuable tools in glycobiology research because they allow scientists to study the effects of glycosylation changes on cell behavior, protein function, and disease processes.

Creation of glycosylation mutants

Gene knockout and gene knockdown are two common techniques used in molecular biology to study the functions of specific genes and their products (proteins) by either eliminating or reducing their expression, respectively. These techniques have various applications, including understanding gene function, disease modeling, and potential therapeutic interventions.

Gene knockout, also known as gene disruption or gene inactivation, is the process of completely eliminating the function of a specific gene in an organism or cell. It involves the permanent removal or disruption of the target gene.

Gene knockdown is valuable for studying gene function when complete gene knockout may not be suitable or necessary. It is used to investigate the role of specific genes in cellular processes, pathways, and diseases. Gene knockdown is commonly used in cell culture experiments and can be employed in highthroughput screening assays to identify potential drug targets. It can be used to assess the impact of reducing gene expression on cellular phenotypes and signaling pathways.

Gene knockdown methods

Gene knockdown is the partial reduction of gene expression, typically by suppressing the transcription or translation of the target gene. Unlike gene knockout, where gene function is completely eliminated, gene knockdown reduces gene expression levels without completely abolishing it.

• In eukaryotic organisms like mice or yeast, gene knockout is often achieved by replacing the target gene with a nonfunctional copy through homologous recombination. This results in the complete loss of gene function.

• The CRISPR-Cas9 system allows for precise gene editing by introducing double-strand breaks in the DNA at the target gene location. This can lead to gene disruption if errors in DNA repair occur.

• In some cases, gene knockout can also be achieved by using RNA interference (RNAi) to degrade the mRNA of the target gene, preventing protein synthesis.

Gene knockout methods

Gene knockout is used to study the function of specific genes by observing the phenotypic changes in organisms or cells lacking that gene. It is valuable for understanding gene involvement in disease development and progression. Gene knockout animals (e.g., knockout mice) are commonly used for modeling human genetic disorders.

• Synthetic Small Interfering RNA (siRNA) molecules are introduced into cells to specifically target the mRNA of the gene of interest. SiRNAs trigger the degradation of the mRNA, leading to reduced protein expression.

• Short Hairpin RNA (shRNA) is similar to siRNA but is expressed from a plasmid or viral vector within the cell, allowing for sustained knockdown over time.

• Antisense Oligonucleotides (ASOs) are short DNA or RNA molecules that bind to the target mRNA, preventing its translation or promoting its degradation.

• CRISPR Interference (CRISPRi) utilizes a modified version of the CRISPR-Cas9 system to inhibit gene expression at the transcriptional level by blocking RNA polymerase binding to the target gene's promoter.

Conclusion

Cultured mammalian cells are indispensable tools in biological and medical research. They offer a controlled and reproducible model system for investigating various aspects of cellular biology, disease mechanisms, and therapeutic development. Careful handling, authentication, and ethical considerations are essential when working with these cells in laboratory settings.Gene knockout and gene knockdown are essential tools in molecular biology and genetics that allow researchers to manipulate gene expression to study gene function and its effects on organisms or cells. The choice between these techniques depends on the specific research goals and the extent of gene manipulation required.