Screening and validation of genome-edited animals.

Screening and validation of genome-edited animals.
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DOI:
10.1177/00236772211016922
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发表时间:
2022-03
期刊:
影响因子:
2.4
通讯作者:
--
中科院分区:
医学4区
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--
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近年来,一系列基因组编辑工具的出现促进了将遗传修饰直接引入胚胎,增加了研究人员在一系列物种中获得等位基因的容易性、效率和目录。除了满足对选择盒的要求并产生除了所需等位基因之外的广泛结果之外,基因组编辑在时间和技术上都改变了等位基因验证过程。而传统的基因靶向依赖于选择,并允许在胚胎干细胞修饰阶段进行等位基因验证,现在筛选预期等位基因的存在发生在(经常是嵌合体)创始动物中。编辑的等位基因的最终确认只能在随后的G1代进行,验证策略必须将所需的等位基因与一系列非预期的结果区分开来。在这里,我们提出了基因编辑带来的一些挑战,验证策略和动物群体管理的考虑因素。
The emergence of an array of genome-editing tools in recent years has facilitated the introduction of genetic modifications directly into the embryo, increasing the ease, efficiency and catalogue of alleles accessible to researchers across a range of species. Bypassing the requirement for a selection cassette and resulting in a broad range of outcomes besides the desired allele, genome editing has altered the allele validation process both temporally and technically. Whereas traditional gene targeting relies upon selection and allows allele validation at the embryonic stem cell modification stage, screening for the presence of the intended allele now occurs in the (frequently mosaic) founder animals. Final confirmation of the edited allele can only take place at the subsequent G1 generation and the validation strategy must differentiate the desired allele from a range of unintended outcomes. Here we present some of the challenges posed by gene editing, strategies for validation and considerations for animal colony management.
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