CRISPR-Cas9 genome editing learning path

CRISPR-Cas9 as genome modification technique

CRISPR-Cas9 is a genome modification (gene-editing) technique that enables scientists to change the DNA of living organisms. It is adapted from a bacterial immune defense system and works by delivering the Cas9 nuclease together with a synthetic guide RNA (gRNA) into cells. The guide RNA directs Cas9 to a specific DNA sequence, where Cas9 cuts both strands of the DNA, allowing researchers to remove existing genes (knock-out) or insert new DNA sequences (knock-in) inside living cells. After Cas9 creates a double-strand break, the cell repairs the DNA using one of two main pathways. Homology-directed repair (HDR) can be used for precise knock-in edits by providing an exogenous repair template with sequences matching the DNA around the cut. Alternatively, non-homologous end joining (NHEJ) or related end-joining pathways such as POLQ/TMEJ often introduce small insertions or deletions, which can disrupt gene function and produce knock-outs. CRISPR-Cas9 is widely valued for being precise, efficient, and cost-effective, with major applications in biotechnology, agriculture, research, and potential treatments for genetic diseases—though germline use in humans remains highly controversial.

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Delivery of Cas9 with guide RNA into cells

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