TY - CHAP
T1 - Molecular aspects of zinc finger nucleases (ZFNs)-mediated gene editing in rat embryos
AU - Cui, Xiaoxia
N1 - Publisher Copyright:
© 2019, Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2019
Y1 - 2019
N2 - 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.
AB - 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.
KW - Conditional knockout
KW - Homology directed repair
KW - Knockin
KW - Knockout
KW - Nonhomologous end joining
KW - Pronuclear microinjection
KW - Single-cell embryos
KW - Zinc finger nucleases
UR - http://www.scopus.com/inward/record.url?scp=85055611845&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-8831-0_17
DO - 10.1007/978-1-4939-8831-0_17
M3 - Chapter
C2 - 30353521
AN - SCOPUS:85055611845
T3 - Methods in Molecular Biology
SP - 295
EP - 312
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -