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Nucleic acids delivery methods for genome editing in zygotes and embryos: the
old, the new, and the old-new
#MMPMID27037013
Sato M
; Ohtsuka M
; Watanabe S
; Gurumurthy CB
Biol Direct
2016[Mar]; 11
(1
): 16
PMID27037013
show ga
In the recent years, sequence-specific nucleases such as ZFNs, TALENs, and
CRISPR/Cas9 have revolutionzed the fields of animal genome editing and
transgenesis. However, these new techniques require microinjection to deliver
nucleic acids into embryos to generate gene-modified animals. Microinjection is a
delicate procedure that requires sophisticated equipment and highly trained and
experienced technicians. Though over a dozen alternate approaches for nucleic
acid delivery into embryos were attempted during the pre-CRISPR era, none of them
became routinely used as microinjection. The addition of CRISPR/Cas9 to the
genome editing toolbox has propelled the search for novel delivery approaches
that can obviate the need for microinjection. Indeed, some groups have recently
developed electroporation-based methods that have the potential to radically
change animal transgenesis. This review provides an overview of the old and new
delivery methods, and discusses various strategies that were attempted during the
last three decades. In addition, several of the methods are re-evaluated with
respect to their suitability to deliver genome editing components, particularly
CRISPR/Cas9, to embryos.
free
free
free
