TY - JOUR T1 - <em>Easi</em>-CRISPR: Efficient germline modification with long ssDNA donors JF - bioRxiv DO - 10.1101/069963 SP - 069963 AU - Rolen M. Quadros AU - Masato Ohtsuka AU - Donald W Harms AU - Tomomi Aida AU - Ronald Redder AU - Hiromi Miura AU - Guy P. Richardson AU - Mark A. Behlke AU - Sarah A. Zeiner AU - Ashley M. Jacobi AU - Lisa D. Urness AU - Suzanne L. Mansour AU - Channabasavaiah B. Gurumurthy Y1 - 2016/01/01 UR - http://biorxiv.org/content/early/2016/08/17/069963.abstract N2 - CRISPR/Cas9 technology efficiently produces short insertions or deletions (indels) and can insert short exogenous sequences at Cas9 cut sites. However, targeting long inserts is still a major technical challenge. To overcome this challenge, we developed Easi-CRISPR (Efficient additions with ssDNA inserts-CRISPR), a method that uses long, in vitro-synthesized, single-stranded DNAs with 50-100 base homology arms as repair templates. We demonstrate that Easi-CRISPR can generate knock-in and floxed alleles in mice with an efficiency at many loci as high as 100%. The simple design requirements for donor DNAs and the reproducibly high-efficiency of Easi-CRISPR enables rapid development of many types of commonly used animal and cell models. ER -