CRISPR Cpf1 Bound to Guide RNA and Target DNA
Explore the 3-dimensional structure of CRISPR Cpf-1 and learn about the future of gene editing. From x-ray crystallography data to 3D printed model, this protein guide will walk you through the inner workings of CRISPR, and ask some really interesting questions about just how unique this protein really is.
“Clustered Regularly Interspaced Short Palindromic Repeats from Prevotella and Francisella CRISPR/Cpf1 is a DNA-editing technology analogous to the CRISPR Cas9 system, characterized in 2015 by Feng Zhang’s group from the Broad Institute and MIT. Cpf1 is an RNA-guided endonuclease of a class II CRISPR/Cas system. This acquired immune mechanism is found in Prevotella and Francisella bacteria. It prevents genetic damage from viruses. Cpf1 genes are associated with the CRISPR locus, coding for an endonuclease that use a guide RNA to find and cleave viral DNA. Cpf1 is a smaller and simpler endonuclease than Cas9, overcoming some of the CRISPR/Cas9 system limitations. CRISPR/Cpf1 could have multiple applications, including treatment of genetic illnesses and degenerative conditions.” https://en.wikipedia.org/wiki/CRISPR/Cpf1
This 3D animation depicts the sturctural properties of the protein pointing out unique properties otherwise hidden from view.
Cpf1 is an RNA-guided endonuclease of a type V CRISPR-Cas system that has been recently harnessed for genome editing. Here, we report the crystal structure of Acidaminococcus sp. Cpf1 (AsCpf1) in complex with the guide RNA and its target DNA at 2.8 Å resolution. AsCpf1 adopts a bilobed architecture, with the RNA-DNA heteroduplex bound inside the central channel. The structural comparison of AsCpf1 with Cas9, a type II CRISPR-Cas nuclease, reveals both striking similarity and major differences, thereby explaining their distinct functionalities. AsCpf1 contains the RuvC domain and a putative novel nuclease domain, which are responsible for cleaving the non-target and target strands, respectively, and for jointly generating staggered DNA double-strand breaks. AsCpf1 recognizes the 5′-TTTN-3′ protospacer adjacent motif by base and shape readout mechanisms. Our findings provide mechanistic insights into RNA-guided DNA cleavage by Cpf1 and establish a framework for rational engineering of the CRISPR-Cpf1 toolbox. “http://www.rcsb.org/pdb/explore.do?structureId=5B43
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This is a 3D print of CRISPR Cpf1 created in full color sandstaone. The model depicts CRISPR (blue) bound to its guide RNA (red) while editing its Target DNA (yellow and green). The subtle color variations in the CRISPR are associated to its bFactor, or atomic temperature. The darker the color, the less the protein moves with a cooler atomic temperature.