“Usage a CRISPR/Cas9-based for obtaining knockouts of economically significant cattle genes”

An experimental work dealing with the gene modification using Cas9 RNA-based editing system was performed. Point site-specific breakpoints in gDNA were introduced at zygote stage by microinjection of spCas9 mRNA protein and guide RNAs into cytoplasm. Oocytes that extruded first second polar bodies used for injection. 2 series microinjections designs early bovine embryos made. The degeneration ranged from 10% to 56% different groups. A total 100 injections Cleavage started 78% surviving oocytes; 5 reached blastocyst stage, which 16% number dividing embryos. All resulting analyzed evaluate efficiency editing. isolated all had stage. Using Sanger sequencing genes interest pre-implantation biopsies them, it shown out 17 RNA against BLG SpCas9 mRNA, 9 after CD209 required genome modifications found. This is indicative high this delivery method system.