Genome Editing with CRISPR-Cas9

What is CRISPR-Cas9 technology? How does CRISPR-Cas9 genome editing work? What is the mechanism of genome engineering with CRISPR-Cas9? What is a PAM sequence? Where is the PAM sequence location and what is its function? What is the protospacer adjacent motif? How does Cas9 recognize target DNA through guide RNA (gRNA) ? How does the Cas9 nuclease cut target DNA? How are the repair mechanisms (HDR, homology-directed repair and NHEJ, Non-homologous end joining) involved in repairing Cas9 double stranded DNA breaks?
This video explains the mechanism of CRISPR-Cas9 in genome editing. Specifically, this video explains how cas9 finds the target DNA match through PAM sequence verification and how the guide RNA pairs with the target DNA. Towards the end, we discuss how multiple repair natural occurring double-stranded DNA break repair pathways like NHEJ (non-homologous end joining) and homology directed repair can be used in CRISPR-Cas9 genome editing, especially when making large deletions or making targeted insertions.

00:00 Cas9 Endonuclease
00:34 single-guide RNA (sgRNA)
01:13 Protospacer adjacent motif (PAM)
02:18 CRISPR-Cas9 Genome editing
04:48 Exercise
05:16 Non-homologous end joining (NHEJ)
06:30 Homology directed repair (HDR)
07:14 Cell cycle and DSB repair
07:54 Cas9 to disrupt the genome
08:12 Cas9 to delete DNA from the genome
08:34 Cas9 directed donor DNA insertion
09:17 CRISPR-Cas9 challenges

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I appreciate your support :)

[brokenpiano]

Thank you! I was looking for details on the orientation of sgRNA/target DNA, your video makes it so simple and clear. Also, I solved the exercise correctly :)

[hurmatarif]

Thank you so much 😊 please make videos on DNA binding domain

[muniraabbaslimkhedawala]

Can you please do cre lox p next please

[somakhaled]

thank you im student biotechcnology from jordan