There are various technologies and techniques currently available for knocking out or editing genes. One of the technologies, which is based on the second approach, is “knockout mice”. Researchers are able to breed mice that have a gene knocked out throughout its entire body. This is achieved by creating a construct of DNA containing a mutated allele, injecting it in cultured embryonic stem (ES) cells, and selecting for the cells that underwent homologous recombination (Figure 1). Then these selected ES cells are injected into early mice embryos, and transferred to a female. If a chimeric mouse is identified a...
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...ort and mucus builds up inside the cell. A study done by Schwank et al. showed they were able to utilize the CRISPR-Cas9 system to repair the CFTR locus. By double stand breaks and homologous recombination in the mutant CFTR gene of the cultured cells of a cystic fibrosis patient, they were able to correct the allele (Figure 4). This was confirmed by PCR and sequencing, and the expressed CFTR protein was observed to be fully functional (5).
CRISPR-Cas9 is a novel technique that is transforming research in biology today. Its advantageous properties compared to knockout mice, and its application in anti-cancer drug testing and genetic engineering prove that it is a potent research tool. Although there are many challenges that lie ahead, this system has great potential for application in other deadly diseases, such as HIV, as well as targeted cancer therapy (1).
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