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Original article link: HeYuan Bio – CRISPR/Cas9
CRISPR/Cas9 is a technology that enables precise editing of specific genomic loci. Its mechanism involves the Cas9 nuclease, guided by a guide RNA (gRNA), which recognizes a target genomic sequence and cleaves double-stranded DNA, generating a double-strand break. Cells then repair this break via non-homologous end joining (NHEJ) or homologous recombination (HR), thereby achieving gene knockout or precise genome editing at the DNA level.
The CRISPR/Cas9 system consists of two main components:
1. Single-guide RNA (sgRNA)
2. Cas9 protein with endonuclease activity
CRISPR gene knockout
When the goal is gene knockout, Cas9–sgRNA can be used to cleave DNA, generating double-strand breaks. The cell then repairs the DNA via non-homologous end joining, during which random insertions or deletions of nucleotides are introduced, resulting in a frameshift mutation that leads to the disruption of the target gene.
CRISPR gene editing
When the goal is gene knock-in or replacement, the principle of homologous recombination must be employed. In the CRISPR/Cas9 system, the Cas9–sgRNA complex induces a double-strand break at the target site; in the presence of a template DNA, the cell repairs the DNA via homologous recombination. The template DNA can be deliberately designed to carry the gene to be inserted or the gene to be repaired, thereby enabling the cell to encode the desired gene.
Selection of the Carrier (Partial)
We offer single- and dual‑vector systems containing Cas9 protein, with viral delivery. Multiple gRNAs can be designed based on the target gene sequence, and the vectors are equipped with various fluorescent and antibiotic markers for convenient selection.
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Carrier Category |
Number |
Carrier element |
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CRISPR lentiviral single vector |
H5070 |
pLenti-U6-spgRNA v2.0-CMV-Puro-P2A-3xFLAG-spCas9-WPRE |
|
H7072 |
pLenti-U6-spgRNA v2.0-CMV-BSR-P2A-3xFLAG-spCas9-WPRE |
|
|
H6825 |
pLenti-U6-spgRNA v2.0-CMV-sfGFP-P2A-3xFLAG-spCas9-WPRE |
|
|
CRISPR lentiviral dual-vector system |
H5068 |
pLenti-U6-spgRNA v2.0-CMV-EGFP-WPRE |
|
H5450 |
pLenti-CMV-Puro-P2A-3xFLAG-espCas9_1.1-WPRE |
|
|
CRISPR AAV single-vector system |
H6941 |
pAAV-CMV-SaCas9-U6-sagRNA v2.0 |
|
H5273 |
pAAV-hSyn-SaCas9-U6-sagRNA v2.0 |
|
|
CRISPR AAV dual-vector system |
H6291 |
pAAV-CMV-EGFP-WPRE-U6-spgRNA v2.0 |
|
H12663 |
pAAV-U6-shRNA/spgRNA v2.0-CMV-EGFP-WPRE |
|
|
H11012 |
pAAV-CMV-Luc2-WPRE-U6-spgRNA |
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