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A conditional gene knockout allows gene deletion in a tissue in a tissue specific manner. This is required in place of a gene knockout if the null mutation would lead to embryonic death, [13] or a specific tissue or cell type is of specific interest. This is done by introducing short sequences called loxP sites around the gene.
The genes are knocked out because this is the best way to study their function and learn more about their role in human diseases. There are two main strategies to conditional gene knockout and those are gene targeting or homologous recombination and gene trapping.
Floxing a gene allows it to be deleted (knocked out), [5] [6] translocated or inserted [7] (through various mechanisms in Cre-Lox recombination). The floxing of genes is essential in the development of scientific model systems as it allows spatial and temporal alteration of gene expression.
Gene knock-in originated as a slight modification of the original knockout technique developed by Martin Evans, Oliver Smithies, and Mario Capecchi.Traditionally, knock-in techniques have relied on homologous recombination to drive targeted gene replacement, although other methods using a transposon-mediated system to insert the target gene have been developed. [3]
Gene knockdown is an experimental technique by which the expression of one or more of an organism's genes is reduced. The reduction can occur either through genetic modification or by treatment with a reagent such as a short DNA or RNA oligonucleotide that has a sequence complementary to either gene or an mRNA transcript.
Vector containing DNA sequence similar to the gene to be modified is introduced to the cell, and by a process of recombination replaces the target gene in the chromosome. This method can be used to introduce a mutation or knock out a gene, for example as used in the production of knockout mice. [29]
In addition, it has been used to engineer stably modified human embryonic stem cell and induced pluripotent stem cell (IPSCs) clones and human erythroid cell lines, [11] [28] to generate knockout C. elegans, [12] knockout rats, [13] knockout mice, [29] and knockout zebrafish. [14] [30] Moreover, the method can be used to generate knockin organisms.
Gene silencing is the regulation of gene ... when genes are knocked out, ... 1,881 miRNAs were in annotated human miRNA loci. miRNAs were predicted to ...