How to make transgenic mice in neurons?

Question

I am confused and curious about the process of making transgenic mice. Can anyone explain and describe in-depth each step of the process one would use to make a transgenic mouse that expresses GFP only in the neurons that produce dopamine (neurotransmitter not protein)? Thank you!

Answer 1

There are multiple steps involved in producing transgenic mice with particular genetic modifications in neurons. Here's a condensed summary:

First, create the transgene:

• Specify the gene or genes that you wish to introduce.

• Choose the right promoters to target specific expression in neurons.

2) Put Together the Transgene:

•Compile the DNA sequence in accordance with your blueprint.

•Incorporate regulatory components such as terminators and promoters.

3) Confirm Transgene:

• Use sequencing to verify the transgene sequence's accuracy.

4) Embryonic mouse microinjection:

•Inject the transgene at the single-cell stage into fertilized mouse embryos.

inject into the fertilize

d egg's pronucleus.

5) implantation of embryos:

•Inject the injected embryos into a surrogate mother mouse's uterus.

6) Check for Progeny with Transgenics:

•To identify transgenic mice, check if the transgene is present.

7) Procreation and Growth:

•Mate the transgenic mice to increase the number of genetically modified mice in the colony.

8) Explain Transgenic Mice's Features:

•Confirm transgene expression in neurons by employing methods such as molecular assays or immunohistochemistry.

9) Keep the Line Alive and Spread It:

• Take good care of the transgenic mouse line in order to support future studies.

•If necessary, provide other researchers access to t

he transgenic line.

Prior to beginning any animal experimentation, it is imperative to adhere to ethical guidelines and secure the necessary approvals. For best outcomes, you should also think about working with a laboratory that has experience working with transgenic mice or speaking with specialists in the field.