CRISPR in Cancer Biology and Therapy

1. CRISPR in Cancer Biology and
Therapy
Omoyayi Ibrahim
20174831@std.neu.edu.tr
Department of Biomedical Engineering
Near East University

2. History of
CRISPR
 CRISPRs were first identified in E. coli in 1987 by a Japanese
scientist, Yoshizumi Ishino, and his team, who accidentally
cloned an unusual series of repeated sequences interspersed
with spacer sequences while analyzing a gene responsible for
the conversion of alkaline phosphatase.
 Emmanuelle Charpentier is the co-inventor of CRISPR. Together
with Dr. Doudna, Dr. Charpentier was involved in the
biochemical characterization of guide RNA and Cas9 enzyme-
mediated DNA cleavage

3. Introduction to
CRISPR Cas 9
System
 CRISPR stands for Clustered Regularly Interspaced
Palindromic Repeats.
 CRISPR-Cas9 was adapted from a naturally occurring
genome editing system that bacteria use as an immune
defense.
 DNA endonuclease that is directed to induce double
strand breaks (DSBs) at specific genomic loci via a
programmable guide RNA (gRNA) molecule that mediates
complementary DNA–RNA base pairing.
 The target sequence must be flanked on the 3′ side by an
‘NGG’ protospacer adjacent motif (PAM) sequence.
 The DSB created by Cas9 can be resolved by either
precise homology-directed repair (HDR) or, non-
homologous end joining (NHEJ)
 HDR enables the introduction of specific changes, while
insertions and deletions (indels) from NHEJ can be
exploited to disrupt coding and noncoding sequences

4. CRISPR Cas 9 System

5. Adaptation of CRISPR / CAS 9 from Prokaryotes into a Therapy in
Eukaryotes
Nature Review, 2020

6. The development of CRISPR tools that can be applied to the study of cancer
biology
Nature Review, 2020

7. Future
Direction on
CRISPR Cas 9
System
Nature Review, 2020

8. References
 1. Sung, H. et al. Global Cancer Statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36
cancers in 185 countries. CA Cancer J. Clin. 71, 209–249 (2021).
 2. Mojica, F. J., Díez-Villaseñor, C., García-Martínez, J. & Soria, E. Intervening sequences of regularly spaced
prokaryotic repeats derive from foreign genetic elements. J. Mol. Evol. 60, 174–182 (2005).
 3. Barrangou, R. et al. CRISPR provides acquired resistance against viruses in prokaryotes. Science 315, 1709–1712
(2007).
 4. Bolotin, A., Quinquis, B., Sorokin, A. & Ehrlich, S. D. Clustered regularly interspaced short palindrome repeats
(CRISPRs) have spacers of extrachromosomal origin. Microbiology 151, 2551–2561 (2005).
 5. Pourcel, C., Salvignol, G. & Vergnaud, G. CRISPR elements in Yersinia pestis acquire new repeats by preferential
uptake of bacteriophage DNA, and provide additional tools for evolutionary studies. Microbiology 151, 653–663
(2005).
 6. Cho, S. W., Kim, S., Kim, J. M. & Kim, J.-S. Targeted genome engineering in human cells with the Cas9 RNA-
guided endonuclease. Nat. Biotechnol. 31, 230–232 (2013).
 7. Jinek, M. et al. RNA-programmed genome editing in human cells. eLife 2, e00471 (2013).
 8. Mali, P. et al. RNA-guided human genome engineering via Cas9. Science 339, 823–826 (2013).
 9. Cong, L. et al. Multiplex genome engineering using CRISPR/Cas systems. Science 339, 819–823 (2013).
 10. Adli, M. The CRISPR tool kit for genome editing and beyond. Nat. Commun. 9, 1911 (2018)

9. Questions?
Feel free to email me any questions you have;
20174831@std.neu.edu.tr