sgRNA Library
Gene knockout is an important method for screening functional loss types and has been widely used in targeted drug research, resistance gene screening, gene function research, genome transcription activation research, protein transport and localization, and other fields.
The target of the widely used RNAi technology is mRNA, while CRISPR recognizes DNA sequences through RNA and then alters them, which can be inherited. Due to the fact that the DNA sequence encoding mRNA only accounts for a very small portion of the total DNA, CRISPR targeting DNA sequences has a much wider range of targets than RNAi, and is more likely to screen for specific CRISPR targets targeting a certain DNA sequence.
In the CRISPR/Cas9 system, the complex of sgRNA and Cas9 endonuclease can generate DNA double strand breaks (DSBs) in the target fragment, inducing frameshift mutations caused by the non homologous terminal junction (NHEJ) repair mechanism. The CRISPR/Cas9 system can simultaneously silence multiple genes and has a wide range of applications, which have been successfully reported in bacteria, yeast, plants, fish, and mammals. At present, more convenient and efficient CRISPR sgRNA libraries have gradually replaced shRNA libraries.
The human gene sgRNA lentivirus library and mouse gene sgRNA AAV library of Weizhen Biotechnology, combined with the stable Cas9 expression cell line provided by Weizhen, will help you achieve high-throughput screening in a shorter time!
一、Weizhen Biological sgRNA Library Related Products and Services
1、sgRNA plasmid/virus
Library Name
|
Target gene name and quantity
|
sgRNA quantity
|
Virus titer
|
Human-derived gene sgRNA lentivirus library
|
Human-derived gene 19114
|
76441
|
1×10^8IU/mL
|
Human-derived gene sgRNA lentivirus library
|
kinase gene 763
|
3052
|
1×10^8IU/mL
|
Mouse -derived gene sgRNA lentivirus library
|
Mouse -derived gene 20611
|
130209
|
1×10^8IU/mL
|
2.Cas9 stable transformation strain
Cell name
|
Source
|
Stably expressed gene
|
HEK293
|
Human
|
Cas9
|
Neuro-2A
|
Mouse
|
Cas9
|
Cell name Source Stably expressed gene
HEK293 Human Cas9
Neuro-2A Mouse
Cell name Source Stably expressed gene
HEK293 Human Cas9
Neuro-2A Mouse
Cell name Source Stably expressed gene
HEK293 Human Cas9
Neuro-2A Mouse
Cell name Source Stably expressed gene
HEK293 Human Cas9
Neuro-2A Mouse
3. Cas9 Stable Transformation Plant Construction Service
Weizhen Biotechnology can construct Cas9 stable transgenic strains based on your experimental needs and the target cells for your experiment. For details, please contact the sales personnel.
二、Activity detection of Cas9 in Weizhen Biotechnology
The activity detection of Cas9 can be achieved by observing the peaks in the sequencing results after transfection of Cas9 plasmid into cells.
Cas9 can cause DNA double strand breaks at recognition sites, which can be easily repaired through error prone repair, leading to random mutations. Therefore, after identifying primers upstream and downstream of the site for PCR, the product is actually a mixture of two types of PCR products, namely the PCR product of the original sequence and the PCR product of the mutant sequence. By sequencing the PCR products, a peak like result will appear, indicating that Cas9 is active. If no peak is observed, it indicates that Cas9 has no activity or extremely low activity.
Cas9 activity detection results (sequencing showed a nested peak result)
三、High throughput screening technology and advantages of sgRNA library
SgRNA library high-throughput screening technology refers to the use of sgRNA plasmid/virus libraries to infect stable Cas9 expressing cell lines, thereby achieving the goal of high-throughput screening.
Compared to traditional shRNA libraries, sgRNA libraries have unparalleled advantages. Not only does it save time and cost in operation, but it also has a wider range of applications and superior decorative effects.
