2. CRISPR……...
…..is complicated.
But we are going to focus on Cas9

5. Who discovered CRISPR?

6. 18 Cell 164, January 14, 2016
342 | NATURE | VOL 535 | 21 JULY 2016

7. Emmanuelle Charpentier
Jennifer
Doudna
Feng Zhang

8. Feng Zhang’s group at MIT.
Where is the boss?
Who are these guys?

9. Emmanuelle Charpentier
Jennifer
Doudna
Emmanuelle Charpentier
Feng Zhang

10. “Finally, the ..history of CRISPR… underscores that scientific breakthroughs are rarely eureka moments. They are typically ensemble acts, played out over a decade or more, in which the cast becomes part of something greater than what any one of them could do alone. It’s a wonderful lesson for the general public, as well as for a young person contemplating a life in science.“
Eric Lander, MIT

11. Who discovered CRISPR? CRISPR discovered? How was
How was CRISPR discovered?
Who discovered CRISPR?
CRISPR discovered?
Flip Words
How
Rearrange
letters
insert
was

13. In the last paragraph of the Discussion…..
This is how science often happens…….

14. …and now a short pause, to consider……..
…… “nyibcviqsofinklmforpkmawerbtics”
…… “nyibcviqsofinklmforpkmawerbtics”
…… “ b i o in for ma tics”
…… “ bioinformatics”
GGAGTTCTACCGCAGAGGCGGGGGAACTCCAAGTGATATCCATCATCGCATCCAGTGCGCCCGGTTT ATCCCCGCTGATGCGGGGAACACCAGCGTCAGGCGTGAAATCTCACCGTCGTTGCCGGTTTATCCCT GCTGGCGCGGGGAACTCTCGGTTCAGGCGTTGCAAACCTGGCTACCGGGCGGTTTATCCCCGCTAAC GCGGGGAACTCGTAGTCCATCATTCCACCTATGTCTGAACTCCCGGTTTATCCCCGCTGGCGCGGGG
Your challenge….

15. Do It………………
….analyze the nucleotide sequence downstream from the iap gene in E. coli….
A paper bioinformatics activity….

16. The nucleotide sequence downstream from the iap gene in E. coli
GGAGTTCTACCGCAGAGGCGGGGGAACTCCAAGTGATATCCATCATCGCATCCAGTGCGCC CGGTTTATCCCCGCTGATGCGGGGAACACCAGCGTCAGGCGTGAAATCTCACCGTCGTTGC CGGTTTATCCCTGCTGGCGCGGGGAACTCTCGGTTCAGGCGTTGCAAACCTGGCTACCGGG CGGTTTATCCCCGCTAACGCGGGGAACTCGTAGTCCATCATTCCACCTATGTCTGAACTCC CGGTTTATCCCCGCTGGCGCGGGGAACTCG
GGAGTTCTACCGCAGAGGCGGGGGAACTCCAAGTGATATCCATCATCGCATCCAGTGCGCCCGGTTT ATCCCCGCTGATGCGGGGAACACCAGCGTCAGGCGTGAAATCTCACCGTCGTTGCCGGTTTATCCCT GCTGGCGCGGGGAACTCTCGGTTCAGGCGTTGCAAACCTGGCTACCGGGCGGTTTATCCCCGCTAAC GCGGGGAACTCGTAGTCCATCATTCCACCTATGTCTGAACTCCCGGTTTATCCCCGCTGGCGCGGGG AACTCG
GGAGTTCTACCGCAGAGGCGGGGGAACTCCAAGTGATATCCATCATCGCATCCAGTGCGCC CGGTTTATCCCCGCTGATGCGGGGAACACCAGCGTCAGGCGTGAAATCTCACCGTCGTTGC CGGTTTATCCCTGCTGGCGCGGGGAACTCTCGGTTCAGGCGTTGCAAACCTGGCTACCGGG CGGTTTATCCCCGCTAACGCGGGGAACTCGTAGTCCATCATTCCACCTATGTCTGAACTCC CGGTTTATCCCCGCTGGCGCGGGGAACTCG

17. CRISPR repeats (29) Spacer sequences (32)
GGAGTTCTACCGCAGAGGCGGGGGAACTCCAAGTGATATCCATCATCGCATCCAGTGCGCC CGGTTTATCCCCGCTGATGCGGGGAACACCAGCGTCAGGCGTGAAATCTCACCGTCGTTGC CGGTTTATCCCTGCTGGCGCGGGGAACTCTCGGTTCAGGCGTTGCAAACCTGGCTACCGGG CGGTTTATCCCCGCTAACGCGGGGAACTCGTAGTCCATCATTCCACCTATGTCTGAACTCC CGGTTTATCCCCGCTGGCGCGGGGAACTCG
Clustered Regularly Interspersed Palindromic Repeats
CRISPR repeats (29)
Spacer sequences (32)
(later shown to be foreign DNA) 29 32

20. What is the PAM? https://youtu.be/iSEEw4Vs_B4
A CRISPR Whiteboard Lesson…presented by the IGI

26. Homology Directed Repair Non- Homologous End Joining
(…think homologous recombination / crossing-over /molecular meiosis )
Homology Directed Repair
Non- Homologous End Joining