Expression of exogenetic enhanced green fluorescent protein in rat endocranium through lentivirus infection.
The study aims to investigate whether exogenetic green fluorescent protein is able to express in the endocranium of rats, and to establish a method for further study in exogenetic gene knock-in or gene overexpression. Forty female Sprague Dawley (SD) rats were randomly divided into 4 groups with 10 in each: low and high dose groups, treated with 10% and 100% EGFP-lentivirus, respectively; negative control group, treated with virus enhancer; sham group, treated with normal saline. Seven days later, half rats' brain tissues were perfusion fixed and fresh brain tissues were obtained from the rest after euthanasia in each group. Immunohistochemical analysis, Western blotting and RT-PCR were respectively performed to detect the site where EGFP expressed and its levels. Immunohistochemical analysis demonstrated that EGFP was successfully expressed in brain tissue of those rats infected with EGFP-lentivirus. Both Western blotting and RT-PCR showed that EGFP was expressed after treatment with EGFP-lentivirus, and the expression level increased with the dosage of the vector. Exogenetic EGFP gene can express in brain tissue of the rat, which laid a solid foundation for future studies in exogenetic gene knock in or gene overexpression.
Department of Neurosurgery, Second Military Medical University, Changhai Hospital Shanghai, China.
Qi
Q
Zhang
Department of Neurosurgery, Second Military Medical University, Changhai Hospital Shanghai, China.
Qiang
Q
Li
Department of Neurosurgery, Second Military Medical University, Changhai Hospital Shanghai, China.
Li
L
Li
Department of Neurosurgery, Second Military Medical University, Changhai Hospital Shanghai, China.
Zhaolong
Z
Zhang
Department of Neurosurgery, Second Military Medical University, Changhai Hospital Shanghai, China.
Yina
Y
Wu
Department of Neurosurgery, Second Military Medical University, Changhai Hospital Shanghai, China.
Yi
Y
Xu
D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation; D055879:Gene Knock-In Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D007150:Immunohistochemistry; D007267:Injections; D016086:Lentivirus / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis; D017207:Rats, Sprague-Dawley; D013997:Time Factors; D014161:Transduction, Genetic / Q000379:methods*
D012333:RNA, Messenger; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
Infection; Lentivirus; enhanced green fluorescent protein (EGFP); exogenetic gene