Production of immunodeficient rabbits by multiplex embryo transfer and multiplex gene targeting.
Production of immunodeficient rabbits by multiplex embryo transfer and multiplex gene targeting.
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DOI：
10.1038/s41598-017-12201-0
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发表时间：
2017-09-22
影响因子：
4.6
通讯作者：
Xu J
中科院分区：
文献类型：
--
作者：
Song J;Yang D;Ruan J;Zhang J;Chen YE;Xu J
免疫缺陷小鼠主要用于生物医学研究。意识到大型动物物种相对于小鼠可能具有增强的预测临床结果的能力，我们致力于通过CRISPR/Cas9开发免疫缺陷兔。我们首先证明了多重胚胎移植有效地产生了多个单基因突变体（SGM）创始人。将用靶向F0 XN 1、RAG 2、IL 2 RG或PRKDC的单一sgRNA显微注射的胚胎合并用于胚胎移植。少至三个受体用于产生四个基因的二十个SGM创始人。然后，我们证明了CRISPR/Cas9强大的多重靶向能力。首先，同时靶向同一染色体上的两个基因，产生三个RAG 1/RAG 2双基因突变体（DGM）创建者。接下来，我们显微注射45个胚胎，每个胚胎具有靶向FOXN 1、RAG 1、RAG 2、IL 2 RG和PRKDC的5种sgRNA，并将它们转移到两个受体中。共产生了5个创始者：1个SGM，2个DGM，1个三基因突变体和1个四基因突变体。目前的工作表明，多重胚胎移植和多重基因打靶可以用来快速，有效地产生突变兔创始人。已经产生了SGM（例如FOXN 1、RAG 2、IL 2 RG和PRKDC）免疫缺陷兔的四个品系以及多基因突变体免疫缺陷兔。这些动物可能被证明对生物医学研究有用。
Immunodeficient mice have been used predominantly in biomedical research. Realizing that large animal species may have an enhanced ability to predict clinical outcome relative to mice, we worked to develop immunodeficient rabbits by CRISPR/Cas9. We first demonstrated that multiplex embryo transfer efficiently produced multiple lines of single-gene mutant (SGM) founders. Embryos microinjected with single sgRNA targeting FOXN1, RAG2, IL2RG or PRKDC were pooled for embryo transfer. As few as three recipients were used to produce twenty SGM founders for four genes. We then demonstrated the powerful multiplex targeting capacity of CRISPR/Cas9. First, two genes on the same chromosome were targeted simultaneously, resulting in three RAG1/RAG2 double-gene mutant (DGM) founders. Next we microinjected forty-five embryos each with five sgRNAs targeting FOXN1, RAG1, RAG2, IL2RG and PRKDC, and transferred them to two recipients. Five founders were produced: one SGM, two DGM, one triple-gene mutant and one quadruple-gene mutant. The present work demonstrates that multiplex embryo transfer and multiplex gene targeting can be used to quickly and efficiently generate mutant rabbit founders. Four lines of SGM (e.g. FOXN1, RAG2, IL2RG, and PRKDC) immunodeficient rabbits, as well as multigenic mutant immunodeficient rabbits have been produced. These animals may prove useful for biomedical research.
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影响因子：
64.5
作者：
Hsu PD;Lander ES;Zhang F
通讯作者：
Zhang F
影响因子：
3.7
作者：
Barbar, Saber-Davide;Pauchard, Laure-Anne;Charles, Pierre-Emmanuel
通讯作者：
Charles, Pierre-Emmanuel
使用CAS9/GRNA系统生成多基因敲除兔子。
DOI：
10.1186/2045-9769-3-12
发表时间：
2014
期刊：
Cell regeneration (London, England)
影响因子：
--
作者：
Yan Q;Zhang Q;Yang H;Zou Q;Tang C;Fan N;Lai L
通讯作者：
Lai L
DOI：
10.1002/path.4648
发表时间：
2016-01
期刊：
The Journal of pathology
影响因子：
--
作者：
Whitelaw CB;Sheets TP;Lillico SG;Telugu BP
通讯作者：
Telugu BP
影响因子：
2.6
作者：
Meier, N;Dear, TN;Boehm, T
通讯作者：
Boehm, T