After You inject your DNA sequence into the distal gonad region using the microinjection, what happens to the DNA? Is it taken up by the germ line nuclei during the developmeant of the oocyte? If so what is the mechanism of this process? eAll of the literature seems to be lacking any details of how the DNA is actually expressed. It just talks about finding the DNA incorperated into the chromosomes or existing as an extrachromosomal DNA array with in the nucleus but does not explain how the DNA makes it’s way into the nucleus in the first place. Is it just randomly taken up by the nucleus?
Im writing a lecture right now on different ways in which you can create transgenic organisms and I want to include microinjecting into the lecture but I just know a student is going to ask me how the DNA is expressed if it’s floating around in the cytoplasm.
Hi, my understanding is that it is taken up by the nucleus when the nucleus is formed during mitosis of an oocyte.
After injection, imagine the series of events as follows: multimerization of injected DNA, possibly driven by homologous sequences, and packaging in DNA packaging proteins present in the cytoplasm. Then, during mitosis, the nuclear envelope is dissolved, and when it is reformed, the extrachromosomal DNA can wind up being within the nucleus and then can be passed on heritably, since it is in a germline cell.
https://www.ncbi.nlm.nih.gov/books/NBK9890/ for mitosis.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC369179/pdf/molcellb00142-0162.pdf and
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC453137/pdf/emboj00110-0380.pdf for further insight.
Please tale a look at this paper:
- Indeed, it has been shown that microinjection of plasmid DNA (pDNA) in the cytoplasm of non-dividing cells resulted in 51% gene expression, while a massive gene expression occurred when the pDNA was injected in the nucleus (1–3).
- In dividing cells the nuclear envelope disassembles on a regular base, which offers an opportunity for DNA to enter the nucleus (4–6).
Especially point 2. There are a few references avaliable mentioned in this paper.
- Badges
- Science method