Highly Efficient Genome Modification of Cultured Primordial Germ Cells with Lentiviral Vectors to Generate Transgenic Songbirds.
Highly Efficient Genome Modification of Cultured Primordial Germ Cells with Lentiviral Vectors to Generate Transgenic Songbirds.
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DOI：
10.1016/j.stemcr.2021.02.015
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发表时间：
2021-04-13
影响因子：
5.9
通讯作者：
Gahr M
中科院分区：
文献类型：
--
作者：
Gessara I;Dittrich F;Hertel M;Hildebrand S;Pfeifer A;Frankl-Vilches C;McGrew M;Gahr M
对生物体进行基因操作的能力使人们对许多物种的功能基因组学有了重要的了解。在鸟类中，基因组的操纵受到单细胞胚胎无法进入的阻碍。在胚胎发育过程中，鸟类原始生殖细胞（PGCs）通过血流迁移并到达性腺原基，在那里它们发育成成熟的生殖细胞。在这里，我们探讨了使用PGCs产生转基因后代的斑胸草雀，这是一个主要的动物模型的性脑分化，声乐学习，声乐交流。从胚胎血液中获得的斑胸草雀PGCs（zfPGCs）在优化的培养基中培养时显着增殖，并保留生殖和干细胞标记的表达。用慢病毒载体转导培养的zfPGC是高效的，导致增强的绿色荧光蛋白的强烈表达。将转导的zfPGC注射到宿主胚胎中，成功地产生了转基因鸣禽。血液来源的斑胸草雀原始生殖细胞（zfPGC）可以在体外扩增慢病毒载体可以有效地在体外扩增zfPGC遗传修饰的zfPGC可以用于产生转基因斑胸草雀在这篇文章中，Gessara和同事使用从胚胎血液中获得的斑胸草雀原始生殖细胞，使用慢病毒载体对鸣禽物种进行遗传修饰。
The ability to genetically manipulate organisms has led to significant insights into functional genomics in many species. In birds, manipulation of the genome is hindered by the inaccessibility of the one-cell embryo. During embryonic development, avian primordial germ cells (PGCs) migrate through the bloodstream and reach the gonadal anlage, where they develop into mature germ cells. Here, we explored the use of PGCs to produce transgenic offspring in the zebra finch, which is a major animal model for sexual brain differentiation, vocal learning, and vocal communication. Zebra finch PGCs (zfPGCs) obtained from embryonic blood significantly proliferated when cultured in an optimized culture medium and conserved the expression of germ and stem cell markers. Transduction of cultured zfPGCs with lentiviral vectors was highly efficient, leading to strong expression of the enhanced green fluorescent protein. Transduced zfPGCs were injected into the host embryo and transgenic songbirds were successfully generated. Blood-derived zebra finch primordial germ cells (zfPGCs) can be expanded in vitro Lentiviral vectors can efficiently transduce zfPGCs in vitro Genetically modified zfPGCs can be used to generate transgenic zebra finches In this article, Gessara and colleagues used zebra finch primordial germ cells obtained from embryonic blood for the genetic modification of a songbird species using lentiviral vectors.
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影响因子：
3.7
作者：
Macdonald J;Glover JD;Taylor L;Sang HM;McGrew MJ
通讯作者：
McGrew MJ
影响因子：
6.5
作者：
Hummel, S;Lynn, EG;Schneider, WJ
通讯作者：
Schneider, WJ
影响因子：
3.7
作者：
Dimitrov L;Pedersen D;Ching KH;Yi H;Collarini EJ;Izquierdo S;van de Lavoir MC;Leighton PA
通讯作者：
Leighton PA
影响因子：
7.7
作者：
Mak SS;Alev C;Nagai H;Wrabel A;Matsuoka Y;Honda A;Sheng G;Ladher RK
通讯作者：
Ladher RK
影响因子：
5.7
作者：
Mooney, Richard
通讯作者：
Mooney, Richard