Efficient Gene Transfer into Neonatal Mouse Brain Using Electroporation

Abstract

In vivo electroporation works as an effective method to transfer exogenous genes into postnatal rodent forebrain. Nevertheless, two deficiencies were found in the reported methods. First, surgical operation brings unnecessary trauma to newborn pups. Second, the procedure was complicated and the transfection efficiency was relatively low. Here we improved the previous electroporation method and make it more simple and efficient. The pulse voltage was decreased to 90 v. DNA injection into one pup's forebrain could be completed within 30 s without any surgical operation. More than 94% of injected neonates survived. Almost 100% of the survivors expressed the introduced gene and the expression persists as long as 20 days after injection. Thus, this method offers a powerful new way for gene function study in postnatal neurogenesis and neural development.

A reliable and flexible gene manipulation strategy in posthatch zebra finch brain

“…We took advantage of a method for gene delivery, in vivo electroporation, to express exogenous DNA constructs in the developing zebra finch brain. Our survivability rate (98%) compares favorably to that obtained for rodents (70–100%); to our knowledge, survivability rates have not been explicitly reported in other birds 34 35 36 . The number of electroporated zebra finch chicks possessing cells expressing FPs (97%) matches, and sometimes exceeds, those achieved for both rodent embryos (70–90%) and developing chickens (<90%) 16 34 36 .…”

Rapid and efficient gene delivery into the adult mouse brain via focal electroporation

“…In vivo electroporation targeting postnatal or adult brains has been reported by several groups 7 8 9 10 11 . Although previous methods provide rapid gene transfer into different types of brain cells, there were still technical problems such as the requirement for higher voltages and specific devices for accurate currency control.…”

“…Currently, electroporation-based gene transfer has been broadly adopted as a convenient gene delivery method for a variety of cell types in developing and adult organs 6 . The electroporation of postnatal and adult brains has also been established in several groups 7 8 9 10 11 . However, high voltage electroporation of whole brains carries the risk of damaging tissues and inhibiting brain functions.…”

Gene Transfer into the Both Sides of Postnatal Cerebral Cortex Using Tweezer Electrode

“…In vivo electroporation works as highly efficient nonviral methods to transfer foreign genes into mouse brain [1][2][3]. Nevertheless, an important limitation of the present in vivo electroporation is that the transfection is unidirectional, and only one side of cerebral cortex can be electroporated [4,5], which may limit its application in gene therapy studies [6,7].…”