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Review
. 2016 Dec 31;4(4):e69.
doi: 10.15190/d.2016.16.

A CRISPR View of Biological Mechanisms

Eduardo Martinez  1 , Lilia Sanchez  1 , Neftali Vazquez  1 , Rebecca Marks  1 , Raechel Cedillo  1 , Christa Respondek  1 , Martin Holguin  1 , Michael W Persans  1 , Megan Keniry  1
Affiliations
Review

A CRISPR View of Biological Mechanisms

Eduardo Martinez et al. Discoveries (Craiova). .

Abstract

A decade ago, only six manuscripts would be found on a PubMed search for "CRISPR," compared to 2,011 manuscripts in 2016. The purpose of this review is to discuss this emergent technology that has revolutionized molecular biological research in just a few years. Endogenous CRISPR mechanisms are harbored by bacteria and archaea as an adaptive defense system that targets foreign DNA from viruses and plasmids. CRISPR has been adapted as a genome editing tool in a plethora of organisms ranging from yeast to humans. This tool has been employed to create loss of function mutations, gain of function mutations, and tagged alleles in a wide range of settings. CRISPR is now extensively employed for genetic screens. CRISPR has also been adapted to study transcriptional regulation. This versatile and relatively facile technique has, and will be, tremendously impactful in research areas such as biomedical sciences, agriculture, and the basic sciences.

Keywords: CRISPR; Cas; Review.

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Conflict of interest statement

Conflict of interests: The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Figures

Figure 1
Figure 1. Unique Genomic Locus noticed in 1987 would later be realized as a CRISPR locus
Nakata et al. discovered repetitive sequences separated by non-repetitive spacer sequences on the 3’ end of the IAP gene in E. Coli in 1987. This striking arrangement of DNA sequences later became known as CRISPR (clustered regularly interspaced short palindromic repeats).
Figure 2
Figure 2. Scheme for Heterologous CRISPR
Schematic demonstrates how the Cas9 exonuclease forms a complex with the synthetic guide RNA (sgRNA) in order to create double strand breaks in a sequence-specific manner. These double strand breaks are resolved by Non Homologous End Joining, which is error-prone commonly leading to insertion or deletions or Homology Directed Repair (HDR) with donor sequences to insert sequences of interest at the break site.
See this image and copyright information in PMC

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