KO cell pool/cell line customization service
What is a KO Cell Pool?
A certain number of wild-type cell lines were transfected by gene editing methods (lentivirus, plasmid method and RNP method, etc.) to obtain a group of successfully edited and unsuccessfully edited cell mixtures.
Why choose KO Cell Pool?
(1) The preparation time of KO Pool is shorter and the cost is lower than that of cell lines, and the results of gene function verification can be obtained quickly;
(2) Genetic diversity: KO Cell Pool usually contains multiple different knockout cells, which is closer to the genetic heterogeneity of the cell population in the natural state. This diversity can provide more comprehensive data to help researchers understand the effects of gene knockouts on cell populations, not just the effects of a single cell line.
(3) Stability of experimental results: Due to the inclusion of a variety of knockout cells, KO Cell Pool has shown higher stability and repeatability in some experiments. In monoclonal cell lines, unintended variation or adaptive changes caused by any gene knockout may cause fluctuations in experimental results, and in the cell pool, this effect may be balanced by diversity.
(4) Suitable for complex biological problems: For the study of complex biological problems, such as multi-gene interactions, signaling pathways, etc., the use of KO Cell Pool may be more appropriate. The diversity ensures that different types of knockout cells can mimic complex biological processes, providing experimental conditions that more closely resemble physiological states.
3. How to obtain the true sense of the KO cell line (KO Cell Line ) ?
(1) KO cell polyclonal cell (KO Cell Pool) is the true KO cell line (KO Cell Line), which is obtained by dilution/flow sorting/monoclonal cell sorting technology, and confirmed by Sanger sequencing that its genotype is the exact fragment loss or frameshift mutation. The KO Pool obtained by lentivirus and plasmid method is a monoclonal clone obtained through drug screening. It can only be determined that the plasmid is integrated into the genome, and it cannot be determined whether the real gene editing occurs. Subsequent experiments are still needed to verify whether the real gene editing occurs.
(2) What is a homozygous KO cell line? All alleles of wild-type cells undergo effective gene editing (non -3-fold frameshift mutation or loss of fragments of more than 21 amino acids), and there is no wild-type gene, which is a homozygous gene knockout cell line. Due to the probability of gene editing, it is difficult to achieve the same editing site of alleles, but there is no wild-type gene, which is a successful homozygote.
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