Techniques to study development Flashcards
What are the 3 general steps when trying to answer a question about animal development?
Find it, move it, restore it
What is a fate map?
A projection onto a developing embryo with what each cell will become
How is a fate map produced?
Tracing cell lineages by tracking cell division and migration using pigments and dyes
What are 4 possible dyes that could be used to trace cell lineages?
- Natural pigments endogenous to the cell
- Vital dyes that diffused into the cell
- Fluorescent dyes
- GFP incorporated into the genome
What is another way to determine a cell lineage without using dyes?
Single cell sequencing to determine the transcriptome of individual cells
What does in situ hybridization tell us?
Where is an mRNA transcript in a cell/embryo being expressed
How could we determine the specific location of a particular protein?
Labelled antibodies
Why can’t antibodies or in situ hybridization be used alone to much effect?
mRNA and proteins can localize differently, two totally separate parts of the central dogma
Once you find the thing you’re interested in in an embryo, what are 4 ways you could move or delete it?
Genetic labelling and transplantation, transgenics, GFP, transplants and hormone manipulation
How do you do genetic labelling and transplantation?
Label some cells from one embryo and transplant them into another embryo that hasn’t been labelled
What is a chimera?
An individual containing genetic information from more than 1 organism
How do you do transgenics?
Recombinant DNA techniques and transfection, mutagenesis, knockdown of gene expression with RNAi, knockout animals. Some way that alters the genetic code to produce a phenotype
How do you reverse the effects of a gene knockdown or knockout?
Treat with bioactive molecules or use transgenic approaches
