CRISPR/Cas9 systems and application of CRISPR/Cas9 systems in construction of alpha, beta and alpha-beta thalassemia model pig cell lines
A thalassemia and cell line technology, applied in the field of gene editing, can solve problems such as difficulty, late sexual maturity, and slow population expansion, achieve good applicability, reduce the generation of non-frameshift mutations, and improve gene editing efficiency. Effect
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment Construction
[0092] 1. Construction of Cas9 high-efficiency expression vector and detection of application effect
[0093] 1.1 Construction of Cas9 high-efficiency expression vector
[0094] The pU6gRNA eEF1a-mNLS-hSpCas9-EGFP-PURO vector (pKG-GE3 for short) was transformed from the addgene (Plasmid#42230, from Zhang Feng lab) pX330-U6-Chimeric_BB-CBh-hSpCas9 vector. Nuclear localization signal, adding WPRE sequence to enhance translation, adding green fluorescence and resistance screening puro gene, the map is as follows figure 1 , the base sequence is shown in SEQ ID NO:70.
[0095] The structure of the original vector pX330-U6-Chimeric_BB-CBh-hSpCas9 used is as follows figure 2 As indicated, purchased from addgene (Plasmid #42230, from Zhang Feng lab).
[0096] The build steps are as follows:
[0097] (1) Remove excess short gRNA backbone
[0098] pX330-U6-Chimeric_BB-CBh-hSpCas9( figure 2 ) was digested with BbsI and XbaI, and the vector fragment (about 8313bp) was recovered, a...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com