Generation of genetically modified rat models via the CRISPR/Cas9 technology
Received date: 2022-11-10
Revised date: 2022-12-23
Online published: 2023-01-01
Supported by
Supported by the National Natural Science Foundation of China(32025023);Supported by the National Natural Science Foundation of China(81873685);Supported by the National Natural Science Foundation of China(31971366);the National Key Research and Development Program of China(2019YFA0110802);the National Key Research and Development Program of China(2019YFA0802802);the National Key Research and Development Program of China(2019YFA0802802);the Shanghai Municipal Commission for Science and Technology(20140900200);the Animal Center of East China Normal University(011)
The RNA-guided CRISPR/Cas9 genomic editing system consists of a single guide RNA (sgRNA) and a Cas9 nuclease. The two components form a complex in cells and target the genomic loci complementary to the sgRNA. The Cas9 nuclease cleaves the target site creating a double stranded DNA break (DSB). In mammalian cells, DSBs are often repaired via error prone non-homologous end joining (NHEJ) or via homology directed repair (HDR) with the presence of donor DNA templates. Micro-injection of the CRISPR/Cas9 system into the rat embryos enables generation of genetically modified rat models. Here, we describe a detailed protocol for creating gene knockout or knockin rat models via the CRISPR/Cas9 technology.
Key words： rat; CRISPR/Cas9; genome editing; gene knockout
Meizhen Liu, Liren Wang, Yongmei Li, Xueyun Ma, Honghui Han, Dali Li . Generation of genetically modified rat models via the CRISPR/Cas9 technology[J]. Hereditas(Beijing), 2023 , 45(1) : 78 -87 . DOI: 10.16288/j.yczz.22-354
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