Biology 1002B Lecture Notes - Lecture 23: Somatic Cell Nuclear Transfer, Human Cloning, Demethylase

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Cloning Primates
Somatic cell nuclear transfer - technique used to clone Dolly
o Before could never clone a monkey
o Take fibroblasts (fetal skin cells) and inject them into donor oocyte that has its
nucleus taken out
o Oocyte thinks that it is fertilized and begins to develop into an embryo using
fibroblast
Epigenetic treatment
o Help fibroblast nucleus get over epigenetic barrier by inserting demethylase so they
do not die earlier
o Monkeys have genomic DNA from their skin cell, so they are clones of the skin
donor
o Monkeys have mitochondrial DNA from oocyte donor
o Monkeys have no DNA from surrogate mother
o Efficiency is still not that good, but we understand epigenetic barrier and know
cloning is possible
Cloning Humans
Therapeutic cloning: make new organs instead of making an entire human
o Take nucleus from a stem cell of a patient and put into an oocyte that is
denucleated
o Get oocyte to grow into embryo
o Take embryonic cells and grow into organs
Human cloning is illegal in Canada
Synthetic Life - create life from scratch
Cells got genome from a computer
These are the first cells that did not come from a pre-existing cell
o Syn1
Typed out genome and put it into empty bacterial cell
The Minimal Genome
1. What is the minimum set of genes needed to support independent cellular life?
o Find organism of smallest genome and see what genes it has
2. How to identify which sequences are essential?
o Find genes that are expressed strongly and common in all cells
o Knock out genes until we discover those that are essential
Genomic Analysis
1. Sequencing
2. Annotation
3. Functional analysis
4. Evolutionary analysis
DNA sequencing methods typically have these basic steps
1. Purification
2. Fragmentation
o Ability to sequence genome only limited to fragments
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