Molecular Aspects of Zinc Finger Nucleases (ZFNs)-Mediated Gene Editing in Rat Embryos.

Molecular Aspects of Zinc Finger Nucleases (ZFNs)-Mediated Gene Editing in Rat Embryos.
Methods Mol Biol. 2019;1874:295-312
Authors: Cui X
Abstract
This chapter contains a collection of protocols involved in using ZFNs to create rat models with various types of genome editing, including simple knockout, point mutation, large deletions, floxing, and insertions. The protocols cover ZFN and donor design criteria, in vitro transcription of ZFNs, validation of ZFNs activity in cultured cells, RNA stability test, microinjection sample preparation, genotyping and in vitro confirmation of floxed alleles, and Southern blot analysis, most of which are not limited to using ZFNs. Instead they apply to model creation in general. When appropriate, a comparison between ZFNs and CRISPR is provided. Standard pronuclear microinjection per se is not discussed.
PMID: 30353521 [PubMed - in process]

Source: Mol Biol Cell - October 27, 2018 Category: Molecular Biology Authors: Cui X Tags: Methods Mol Biol Source Type: research