Skip to main content

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.