Protocol for the electroporation of CRISPR-Cas for DNA and RNA targeting in Bos taurus zygotes.
| Autor: | Biase FH; Virginia Polytechnique Institute and State University, Blacksburg, VA 24061, USA. Electronic address: fbiase@vt.edu., Schettini G; Virginia Polytechnique Institute and State University, Blacksburg, VA 24061, USA. |
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| Jazyk: | angličtina |
| Zdroj: | STAR protocols [STAR Protoc] 2024 Mar 15; Vol. 5 (1), pp. 102940. Date of Electronic Publication: 2024 Mar 08. |
| DOI: | 10.1016/j.xpro.2024.102940 |
| Abstrakt: | The use of CRISPR-Cas9 ribonucleoproteins has revolutionized manipulation of genomes. Here, we present a protocol for the electroporation of CRISPR-Cas for DNA and RNA targeting in Bos taurus zygotes. First, we describe steps for production and preparation of presumptive zygotes for electroporation. The first electroporation introduces ribonucleoproteins formed by Cas9D10A with two guide RNAs to target DNA, and the second introduces the same ribonucleoprotein complex to target DNA plus Cas13a with one guide RNA to target RNAs. For complete details on the use and execution of this protocol, please refer to Nix et al. 1 . Competing Interests: Declaration of interests F.H.B. has a provisional patent application related to the method described in this paper (U.S. Provisional Application No. 63/538,181). (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.) |
| Databáze: | MEDLINE |
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