1. Next Assignment
Presentation on genome editing
Crispr- associated nucleases
Homing endonucleases
Zinc-finger nucleases
Transcription activator-like effector nucleases (TALENs)
Triple-helix–forming oligonucleotides conjugated to restriction endonucleases
CRE/LOX system

2. Epigenetics
heritable chromatin modifications are associated with activated & repressed genes

3. Generating methylated DNA
Si RNA are key: RNA Pol IV generates antisense or foldback RNA, often from TE

4. Generating methylated DNA
Si RNA are key: RNA Pol IV generates antisense or foldback RNA, often from TE
RDR2 makes it DS, 24 nt siRNA are generated by DCL3

5. Generating methylated DNA
RDR2 makes it DS, 24 nt siRNA are generated by DCL3
AGO4 binds siRNA, complex binds target & Pol V

6. Generating methylated DNA
RDR2 makes it DS, 24 nt siRNA are generated by DCL3
AGO4 binds siRNA, complex binds target & Pol V
Pol V makes intergenic RNA, associates with AGO4-siRNA to recruit “silencing Complex” to target site

7. Generating methylated DNA
RDR2 makes it DS, 24 nt siRNA are generated by DCL3
AGO4 binds siRNA, complex binds target & Pol V
Pol V makes intergenic RNA, associates with AGO4-siRNA to recruit “silencing Complex” to target site
Amplifies signal!
extends meth-
ylated region

8. Cytoplasmic regulation
lifetime
localization
initiation

9. Post-transcriptional regulation
Nearly ½ of human genome is transcribed, only 1% is CDS
98% of RNA made is non-coding

10. Post-transcriptional regulation
Nearly ½ of human genome is transcribed, only 1% is CDS
98% of RNA made is non-coding
~1/3 intron

11. Post-transcriptional regulation
Nearly ½ of human genome is transcribed, only 1% is CDS
98% of RNA made is non-coding
~1/3 intron
~2/3 “independently transcribed”