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CRISPR/Cas9 Gene Knockout Technology (CAT#: STEM-MB-0081-WXH)

Introduction

Clustered regularly interspaced short palindromic repeats (CRISPR), is the the DNA that makes up the bacterial adaptive immune system. It is used to help protect bacteria against invading viruses and facilitate their destruction by providing a memory of target DNA sequences to cut. These repeating spacer sequences guide nucleases to cut DNA of the same sequence that invades the cell in the future. This DNA cleavage is achieved by the enzyme Cas9, which has the ability to cut through DNA.
Gene knockout is divided into two types: complete gene knockout and conditional gene knockout: complete gene knockout refers to the complete elimination of target gene activity in cells or individual animals through gene knockout technology. Conditional gene knockout refers to gene knockout at a specific time and space through a positional recombination system.
Technical advantages:
* High editing efficiency, simple construction and convenient operation
* Strict screening criteria can more effectively remove false positives
* low cost
* Broad-spectrum, no gene, cell and species restrictions
* Simultaneous gene targeting of multiple target sites can be achieved
* Rich in functions, can realize multiple target genes such as knockout, insertion, suppression, and activation




Applications

• Scientific research such as genotyping, epidemiological research, analysis of bacterial flora differences in different human bodies, and detection of phages in the environment.
• Construction of phage-resistant industrial production strains;
• As a tool for genetic manipulation, for targeted gene silencing
• Site-specific cleavage of DNA and RNA molecules in vitro

Procedure

1. gRNA design
2. Cell Transfection
3. Single cell cloning
4. Monoclonal Screening
5. Monoclonal Sequencing Analysis

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