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Table 1 Nucleic acids delivery methods in animal transgenesis and genome editing

From: Nucleic acids delivery methods for genome editing in zygotes and embryos: the old, the new, and the old-new

Category Method Remarks Germ line transmission potential
Ex vivo approaches Pronuclear injection The most commonly used method followed by thousands of labs for over 3 decades High
Viral Vectors A few labs used. Limited success. High when lentiviral vectrs are used
Receptor-mediated uptake Only one report [28]. Not proven
In vitro electroporation Novel approach: also proven using CRISPR system. High
Liposomal transfection Very few labs used. Limited success. Not proven
Blastocyst microinjection Only one report [32]: may be suitable for expression analysis in embryonic tissues. Not proven
Sperm-mediated gene transfer (SMGT) Very few labs have attempted. Limited success. Low
Intracytoplasmic sperm injection-mediated gene transfer (ICSI-MGT) Very few labs have attempted. Limited success. Low
In vivo delivery to pre-implantation embryos, fetuses and ovarian tissues GONAD Only one report [50]. This method completely eliminates the need for isolation, microinjection and transfer of embryos to recipient mice. Only one recent so far, yet to be tested in other labs. Not proven yet, but highly likely
Trans-placental gene delivery to fetuses Very few labs have attempted. Limited success. Very low
Delivery to fetal tissues in utero Very few labs have attempted. Limited success. Very low
In vivo delivery to ovarian tissues Very few labs have attempted. Limited success. Low
In vivo delivery to male gonadal tissues Testis-mediated gene transfer (TMGT) Several labs have attempted. Limited success. Possible, may need to screen many offspring from the treated males
Seminiferous tubule-mediated gene delivery A few labs have attempted. Limited success.
Gene delivery via vas deferens Very few labs have attempted. Limited success.
Nucleic acids delivery to the cauda epididymis Very few labs have attempted. Limited success.