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Near-infrared fluorescent protein iRFP713 as a reporter protein for optogenetic vectors, a transgenic Cre-reporter rat, and other neuronal studies.近红外荧光蛋白iRFP713作为光遗传学载体、转基因Cre报告大鼠及其他神经元研究的报告蛋白。
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Efficient gene targeting in mouse zygotes mediated by CRISPR/Cas9-protein.由CRISPR/Cas9蛋白介导的小鼠受精卵高效基因靶向。
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Targeted Germline Modifications in Rats Using CRISPR/Cas9 and Spermatogonial Stem Cells.利用CRISPR/Cas9和精原干细胞对大鼠进行靶向生殖系修饰。
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Disease pathways at the Rat Genome Database Pathway Portal: genes in context-a network approach to understanding the molecular mechanisms of disease.疾病通路在大鼠基因组数据库通路门户：基因背景下的网络方法理解疾病的分子机制。
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Somatic mosaicism and allele complexity induced by CRISPR/Cas9 RNA injections in mouse zygotes.CRISPR/Cas9 RNA注射诱导小鼠受精卵中的体细胞嵌合现象和等位基因复杂性。
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One-step generation of mice carrying reporter and conditional alleles by CRISPR/Cas-mediated genome engineering.通过 CRISPR/Cas 介导的基因组工程一步生成携带报告基因和条件性等位基因的小鼠。
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Heritable gene targeting in the mouse and rat using a CRISPR-Cas system.利用CRISPR-Cas系统在小鼠和大鼠中进行可遗传的基因靶向。
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Genome sequencing reveals loci under artificial selection that underlie disease phenotypes in the laboratory rat.基因组测序揭示了人工选择下的基因座，这些基因座是实验室大鼠疾病表型的基础。
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长 Evans 大鼠精原细胞系是用于转基因大鼠生产的有效种系载体。

Long Evans rat spermatogonial lines are effective germline vectors for transgenic rat production.

作者信息

Hamra F Kent, Richie Christopher T, Harvey Brandon K

机构信息

Department of Pharmacology, University of Texas Southwestern Medical Center, 6001 Forest Park Road, Dallas, TX, 75390, USA.

Department of Obstetrics and Gynecology, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, TX, 75390, USA.

出版信息

Transgenic Res. 2017 Aug;26(4):477-489. doi: 10.1007/s11248-017-0025-2. Epub 2017 Jun 12.

DOI:10.1007/s11248-017-0025-2
PMID:28608322
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7336504/
Abstract

Long Evans rat strains are applied as research models in a broad spectrum of biomedical fields (>15,800 citations, NCBI PubMed). Here, we report an approach to genetically modify the Long Evans rat germline in donor spermatogonial stem cells. Long Evans rat spermatogonial lines were derived from freshly isolated laminin-binding spermatogonia. Laminin-binding spermatogonia were cultured over multiple passages on fibroblast feeder layers in serum-free culture medium containing GDNF and FGF2. Long Evans rat spermatogonial lines were genetically modified by transposon transduction to express a germline, tdTomato reporter gene. Donor rat spermatogonial lines robustly regenerated spermatogenesis after transplantation into testes of busulfan-treated, allogenic, Long Evans rats. Donor-derived spermatogenesis largely restored testis size in the chemically sterilized, recipient Long Evans rats. Recipient Long Evans rats stably transmitted the tdTomato germline marker to subsequent generations. Overall, Long Evans rat spermatogonial lines provided effective donor germline vectors for genetically modifying Long Evans rats.

摘要

长 Evans 大鼠品系被用作广泛生物医学领域的研究模型（>15,800 次引用，NCBI PubMed）。在此，我们报告一种在供体精原干细胞中对长 Evans 大鼠种系进行基因改造的方法。长 Evans 大鼠精原细胞系源自新鲜分离的层粘连蛋白结合精原细胞。层粘连蛋白结合精原细胞在含有 GDNF 和 FGF2 的无血清培养基中，在成纤维细胞饲养层上进行多次传代培养。长 Evans 大鼠精原细胞系通过转座子转导进行基因改造，以表达种系 tdTomato 报告基因。供体大鼠精原细胞系在移植到经白消安处理的同种异体、长 Evans 大鼠的睾丸后，有力地再生了精子发生。供体来源的精子发生在化学绝育的受体长 Evans 大鼠中很大程度上恢复了睾丸大小。受体长 Evans 大鼠将 tdTomato 种系标记稳定地传递给后代。总体而言，长 Evans 大鼠精原细胞系为基因改造长 Evans 大鼠提供了有效的供体种系载体。