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Crispr/cas9 applications, limits and drawbacks

Lusseveld, Jarnick (2019) Crispr/cas9 applications, limits and drawbacks. Master's Thesis / Essay, Molecular Biology and Biotechnology (2016-2019).


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Discovery of the CRISPR/Cas9 system is a promising breakthrough in genome editing, because it can make specific gene alterations more quickly and efficiently than previews techniques (Christopher A. Lino, Jason C. Harper, 2018; Selle & Barrangou, 2015). The CRISPR/Cas system can introduce double stranded breaks (DSB) at specific sites (Ceasar, Rajan, Prykhozhij, Berman, & Ignacimuthu, 2016). These DSBs can be repaired by the non-homologous end joining pathway (NHEJ) and homologous directed repair pathway (HDR). NHEJ will result in random inserts or deletions (Bialk, Rivera-Torres, Strouse, & Kmiec, 2015), whereas HDR repairs DSBs by use of a homologous DNA sequence. HDR can be used for specific genome editing, such as knock-ins or knock-outs. The CRISPR/Cas9 system consists of three components, the Cas9 protein, the guide RNA (sgRNA), and a single stranded oligodeoxynucliotides (ssODN) which provides the homologous sequence carrying the mutation. CRISPR/Cas9 has been used extensively for many applications to great success (Maruyama et al., 2015; Tang & Liu, 2018; Yi & Li, 2016). In this essay, applications, limitations and the potential to overcome these limitations are discussed. CRISPR/Cas9 does have some limitations as genome editing tool. For example, several mismatches are allowed in the guide RNA as well in the PAM site, increasing the change of off-target DSBs. CRISPR/Cas9 has great potential to correct gene alterations in humans, however careful consideration of limitations is required and several hurdles need to be overcome for usage in therapeutic studies (Oude Blenke, Evers, Mastrobattista, & van der Oost, 2016).

Item Type: Thesis (Master's Thesis / Essay)
Supervisor name: Haastert, P.J.M. van
Degree programme: Molecular Biology and Biotechnology (2016-2019)
Thesis type: Master's Thesis / Essay
Language: English
Date Deposited: 08 Jan 2019
Last Modified: 18 Jan 2019 14:40

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