Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other...
-
Upload
blanche-osborne -
Category
Documents
-
view
216 -
download
1
Transcript of Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other...
![Page 1: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/1.jpg)
Final Journal ClubMonday April 27 & Wed April 291.New techniques for genome editing & other purposes• CRISPR• TALEN• Zn Finger• Cre-Lox
2.New techniques for DNA sequencing• Illumina• Ion Torrent• Nano-Pore• Pac-Bio• Mass-Spec• Other?
3.Molecular biology for fun and profit
![Page 2: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/2.jpg)
Telomeres• sequences at chromosome ends • humans have 250-7,000 repeats of TTAGGG• special proteins bind them
![Page 3: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/3.jpg)
Telomeres• sequences at chromosome ends • humans have 250-7,000 repeats of CCCTAA• special proteins bind them• can't replicate 5' end of lagging strand since remove primer
![Page 4: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/4.jpg)
Telomeres• can't replicate 5' end of lagging strand since remove primer• telomeres lose ~ 200 bp each S
![Page 5: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/5.jpg)
Telomeres• can't replicate 5' end of lagging strand since remove primer• telomeres lose ~ 200 bp each S• telomerase replaces missing bases
![Page 6: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/6.jpg)
Telomerase• telomeres lose ~ 200 bp each S• telomerase replaces missing basesreverse transcriptase with attached RNA template
![Page 7: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/7.jpg)
Telomerase• telomeres lose ~ 200 bp each S• telomerase replaces missing basesreverse transcriptase with attached RNA template1) RNA bonds leading strand
![Page 8: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/8.jpg)
Telomerasereverse transcriptase with attached RNA template1) RNA bonds leading strand2) Forms template to extend leading strand
![Page 9: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/9.jpg)
Telomerasereverse transcriptase with attached RNA template1) RNA bonds leading strand2) Forms template to extend leading strand3) Translocates6 bases & repeats
![Page 10: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/10.jpg)
Telomerase1) RNA bonds leading strand2) Forms template to extend leading strand3) Translocates6 bases & repeats4) Extend lagging strandwith primer & DNA pol
![Page 11: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/11.jpg)
Telomere structureBound by complex called shelterin or the telosome
![Page 12: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/12.jpg)
Telomere structureBound by complex called shelterin or the telosomeProtects DNA, prevents recombination
![Page 13: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/13.jpg)
Telomere structureBound by Shelterin/telosome complexProtects DNA, prevents recombinationDNA proposed to form T-loop structure IF long enough
![Page 14: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/14.jpg)
Telomere structureBound by Shelterin/telosome complexProtects DNA, prevents recombinationDNA proposed to form T-loop structure IF long enoughG-rich SS-DNA invades and displacesC-rich strand
![Page 15: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/15.jpg)
Telomere structureBound by Shelterin/telosome complexProtects DNA, prevents recombinationDNA proposed to form T-loop structure IF long enoughG-rich SS-DNA invades and displacesC-rich strand POT1/TPP1 binds & protects D-loop
![Page 16: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/16.jpg)
Telomere structureBound by Shelterin/telosome complexProtects DNA, prevents recombinationDNA proposed to form T-loop structure IF long enoughG-rich SS-DNA invades and displacesC-rich strand POT1/TPP1 binds & protects D-loop If can’t form T-loop trigger apoptosis
![Page 17: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/17.jpg)
Telomere Length MaintenanceBound by shelterin/telosome complex Proposed to form T-loop structure IF long enoughIf can’t form T-loop trigger apoptosisTelomere length proposed to be regulated by TRF1 & PINX1
![Page 18: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/18.jpg)
Telomere Length MaintenanceTelomere length proposed to be regulated by TRF1 & PINX1 If too short, not enough TRF1bind so telomerase is active
![Page 19: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/19.jpg)
Telomere Length MaintenanceTelomere length proposed to be regulated by TRF1 & PINX1 If too short, not enough TRF1bind so telomerase is activeOnce long enough TRF1 bindstelomere
![Page 20: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/20.jpg)
Telomere Length MaintenanceTelomere length proposed to be regulated by TRF1 & PINX1 If too short, not enough TRF1bind so telomerase is activeOnce long enough TRF1 bindstelomere PINX1 then binds TRF1 & stops telomerase
![Page 21: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/21.jpg)
Telomeres
Aging theory:” mature” cells lose telomerase
lose DNA each S
![Page 22: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/22.jpg)
Telomeres
Aging theory:” mature” cells lose telomerase
lose DNA each S
Die when lose too much: Hayflick limit ~50 divisions for 1˚ cultures
![Page 23: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/23.jpg)
Telomeres
Aging theory:” mature” cells lose telomerase
lose DNA each S
Die when lose too much: Hayflick limit ~50 divisions for 1˚ cultures
Ramunas et al (2015) DOI: 10.1096/fj.14-259531 found that transient expression of TERT in cultured cells added 1000 bp to telomeres and allowed 28 x more divisions in fibroblasts
![Page 24: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/24.jpg)
Telomeres
Aging theory:” mature” cells lose telomerase
lose DNA each S
Die when lose too much: Hayflick limit ~50 divisions for 1˚ cultures
Ramunas et al (2015) DOI: 10.1096/fj.14-259531 found that transient expression of TERT in cultured cells added 1000 bp to telomeres and allowed 28 x more divisions in fibroblasts
Cancer cells must maintain telomere length
•90% reactivate telomerase
![Page 25: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/25.jpg)
Telomeres
Cancer cells must maintain telomere length
•90% reactivate telomerase
• [serum telomerase] can diagnose cancer
![Page 26: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/26.jpg)
Telomeres
Cancer cells must maintain telomere length
•90% reactivate telomerase
• [serum telomerase] can diagnose cancer
• Inhibiting telomerase kills most cancer cells
![Page 27: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/27.jpg)
Telomeres
Cancer cells must maintain telomere length
•90% reactivate telomerase
• [serum telomerase] can diagnose cancer
• Inhibiting telomerase kills most cancer cells
• Telomerase is symptom cf cause of cancer
• not viewed as good target for cancer therapy
![Page 28: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/28.jpg)
Telomeres90% of cancer cells reactivate telomerase
[serum telomerase] can diagnose cancerInhibiting telomerase kills most cancer cellsTelomerase is symptom cf cause of cancer
10 % of cancer cells lengthen telomeres by alternative mechanism involving homologous recombination
![Page 29: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/29.jpg)
Telomeres10 % of cancer cells lengthen telomeres by alternative mechanism involving homologous recombination•normally inactive: therefore good target for cancer therapy
![Page 30: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/30.jpg)
Telomeres10 % of cancer cells lengthen telomeres by alternative mechanism involving homologous recombination•normally inactive: therefore good target for cancer therapy•See high frequencyof SCE at telomeres in ALT cells
![Page 31: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/31.jpg)
Telomeres10 % of cancer cells lengthen telomeres by alternative mechanism involving homologous recombination•normally inactive: therefore good target for cancer therapy•See high frequency of SCE at telomeres in ALT cells•Also see high frequency of telomere fragments• Substrates for HR and telomere elongation
![Page 32: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/32.jpg)
Telomeres10 % of cancer cells lengthen telomeres by alternative mechanism involving homologous recombination•normally inactive: therefore good target for cancer therapy•See high frequency of SCE at telomeres in ALT cells•Also see high frequency of telomere fragments• Substrates for HR and telomere elongation
Key common feature = chromatin remodeling allowing HR•Perhaps due to depletion of shelterin, especially TRF2
![Page 33: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/33.jpg)
DNA damageDNA gets damaged a lot!
![Page 34: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/34.jpg)
DNA damageDNA gets damaged a lot!
![Page 35: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/35.jpg)
DNA damageDNA gets damaged a lot! >200,000 events/human cell/day
![Page 36: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/36.jpg)
DNA damageOccurs 2 ways
1) spontaneously
![Page 37: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/37.jpg)
DNA damageOccurs 2 ways
1) spontaneously2) mutagens : damage DNA
![Page 38: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/38.jpg)
Spontaneous mutations1) In Sa) mispairsC tautomersbind A T tautomersbind G
![Page 39: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/39.jpg)
Spontaneous mutations1) In Sa) mispairsC tautomersbind A T tautomersbind G1/106
![Page 40: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/40.jpg)
Spontaneous mutations1) In Sa) mispairsC tautomersbind A T tautomersbind G1/106
6000/S
![Page 41: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/41.jpg)
Spontaneous mutationsWobbling: no numbers!
![Page 42: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/42.jpg)
Spontaneous mutations1) In Sa) mispairsb) misaligned bases
![Page 43: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/43.jpg)
Spontaneous mutations1) In Sa) mispairsb) misaligned bases
-> indels
![Page 44: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/44.jpg)
Spontaneous mutations1) In S
a) mispairsb) misaligned bases -> indels
2) Hydrolysisa) C deaminates to U
![Page 45: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/45.jpg)
Spontaneous mutations2) Hydrolysis
a) C deaminates to U100-500/cell/day
![Page 46: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/46.jpg)
Spontaneous mutations2) Hydrolysis
a) C deaminates to Ub) 5-meC deaminates to T!
![Page 47: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/47.jpg)
Spontaneous mutations2) Hydrolysis
a) C deaminates to Ub) 5-meC deaminates to T!c) Adenine deaminates to hypoxanthine: bonds C!
![Page 48: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/48.jpg)
Spontaneous mutations2) Hydrolysis
a) C deaminates to U100-500/cell/dayb) Depurination (lose A or G) (also depyrimidation, but 20 x less)
![Page 49: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/49.jpg)
Spontaneous mutations2) decomposition
a) C deaminates to U100-500/cell/dayb) Depurination (lose A or G) (also depyrimidation, but 20 x less)Replace missing base randomly
![Page 50: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/50.jpg)
decompositiona) C deaminates to U 100-500/cell/dayb) Depurination (lose A or G) (also depyrimidation, but 20 x less)Replace missing base randomly 10,000 – 50,000 depurinations /cell / day1,000 – 3,000 depyrimidations /cell / day
![Page 51: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/51.jpg)
Spontaneous mutations2) decomposition3) Oxidation: > 20 base lesions due to oxidation have been identified
![Page 52: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/52.jpg)
Spontaneous mutations2) decomposition3) Oxidation: > 20 base lesions due to oxidation have been identified•G is most reactive: most frequent lesion is 8-hydroxy G
![Page 53: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/53.jpg)
Spontaneous mutations2) decomposition3) Oxidation: > 20 base lesions due to oxidation have been identified•G is most reactive: most frequent lesion is 8-hydroxy G•8oxoG (syn) bonds A well enough to evade proofreading!
![Page 54: Final Journal Club Monday April 27 & Wed April 29 1.New techniques for genome editing & other purposes CRISPR TALEN Zn Finger Cre-Lox 2.New techniques.](https://reader030.fdocuments.us/reader030/viewer/2022032607/56649ed05503460f94bde987/html5/thumbnails/54.jpg)
Spontaneous mutations3) Oxidation: > 20 base lesions due to oxidation have been identified•G is most reactive: most frequent lesion is 8-hydroxy G•8oxoG (syn) bonds A well enough to evade proofreading! •(8oxoG (anti) bonded to C gets removed!)