TY  - GEN
T1  - CRISPRcruncher: A tool for engineering restriction sites into coding regions
AU  - Fay, Samuel F.
AU  - Fay, David S.
AU  - Chhatre, Vikram E.
DO  - 10.17912/micropub.biology.000343
UR  - http://beta.micropublication.org/journals/biology/micropub-biology-000343/
AB  - The precise engineering of genomes using CRISPR/Cas9 homology-directed repair has become largely routine, particularly for making small changes such as nucleotide substitutions (Dickinson and Goldstein, 2016; Farboud, 2017; Sternberg and Doudna, 2015). This approach is often accompanied by the introduction of a restriction endonuclease (RE) site positioned adjacent to the modified target region (Friedland et al., 2013; Lo et al., 2013; Paix et al., 2015; Waaijers et al., 2013; Zhao et al., 2014). Such RE sites allow for rapid PCR-based screening to detect cells or organisms containing the desired edit and greatly aid in subsequent genotyping. In addition, the precise location of the engineered RE site relative to the protospacer adjacent motif (PAM) site and the region targeted for modification can impact the efficiency of CRISPR (Farboud et al., 2019). Currently, most labs design these introduced RE sites without the use of a computational tool (i.e., ‘by hand’), which is inefficient and likely to overlook a large majority of engineerable sites.
PY  - 2021
JO  - microPublication Biology
ER  -