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Advances in tools for designing guide RNAs and repair templates for efficient CRISPR-Cas9-induced homology-directed repair
Poster Title: Advances in tools for designing guide RNAs and repair templates for efficient CRISPR-Cas9-induced homology-directed repair
Submitted on 25 Oct 2017
Author(s): Steven R. Lenger, John A. Schiel, Maren Mayer, Chris Edwards, John Quinn, Shawn McClelland, Melissa L. Kelley, Annaleen Vermeulen, Anja van Brabant Smith
Affiliations: Dharmacon part of Horizon Discovery Group
This poster was presented at GDI conference 2017
Poster Views: 815
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Poster Information
Abstract: Combining these advances with our experimental observations has enabled the development of three tools that significantly speed up and simplify the process of 1) selecting appropriate CRISPR RNAs (crRNAs), 2) designing single-stranded DNA oligo donors (ssDNA), and 3) generating plasmid DNA donor templates to enable scientists to leverage the power of HDR in their research. Successful HDR applications begin with the selection of crRNAs near a desired genomic-sequence alteration site and requires high functionality and specificity.The Dharmacon CRISPR Design Tool incorporates both a functional algorithm for evaluating DNA cleavage efficacy and a novel approach for rapidly and thoroughly evaluating guide RNA specificity. The Edit-R HDR Donor Designer enables a researcher to select the specific bases to be either removed or inserted, define the length of each donor homology arm, and automatically creates silent mutations within the donor template to ensure that the genomic target site of HDR-based editing will not be susceptible to further cleavage by Cas9. Similarly, the plasmid donor tool enables researchers to design the necessary components for rapid assembly of donor plasmids for insertion of longer sequences, such as a fluorescent reporter, and automatically determines which silent mutations would be most appropriate for the crRNA being employed.Summary: Recent studies have resulted in elucidation of many of the key requirements for achieving precise CRISPR-Cas9 gene engineering using homology-directed repair (HDR).Report abuse »
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