02176naa a2200349 a 450000100080000000500110000800800410001902400380006010000140009824501290011226000090024152011840025065000110143465000100144565000240145565300160147965300190149565300100151465300130152465300200153765300190155765300090157665300100158565300200159570000150161570000150163070000160164570000170166170000130167870000170169177301180170810634142022-07-12       2022    bl uuuu     u00u1 u    #d7 a10.1007/978-1-0716-2301-5_162DOI1 aKALDS, P.  aGeneration of Double-Muscled Sheep and Goats by CRISPR /Cas9-Mediated Knockout of the Myostatin Gene.h[electronic resource]  c2022  aAbstract: The myostatin (MSTN) gene has shown to play a critical role in the regulation of skeletal muscle mass, and the translational inhibition of this gene has shown increased muscle mass, generating what is known as ?double-muscling phenotype.? Disruption of the MSTN gene expression using the CRISPR/Cas9 genome-editing system has shown improved muscle development and growth rates in livestock species, including sheep and goats. Here, we describe procedures for the generation of MSTN knockout sheep and goats using the microinjection approach of the CRISPR/Cas9 system, including the selection of targeting sgRNAs, the construction of CRISPR/Cas9 targeting vector, the in vitro examination of system efficiency, the in vivo targeting to generate MSTN knockout founders, the genomic and phenotypic characterization of the generated offspring, and the assessment of off-target effects in gene-edited founders through targeted validation of predicted off-target sites, as well as genome-wide off-target analysis by whole-genome sequencing. Editing the MSTN gene using the CRISPR/Cas9 system might be a rapid and promising alternative to promote meat production in livestock.  aCABRAS  aOVEJA  aPRODUCCION DE CARNE  aCRISPR/Cas9  aGenome editing  aGoats  aKnockout  aMeat production  aMicroinjection  aMSTN  aSheep  aSmall ruminants1 aCRISPO, M.1 aTESSON, L.1 aANEGÓN, I.1 aCHEN KEY, Y.1 aWANG, X.1 aMENCHACA, A.  tMethods in Molecular Biology, 2022, Volume 2495, Pages 295-323. Doi: https://doi.org/10.1007/978-1-0716-2301-5_16