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Case Studies

• Experience with Base Editor and Prime Editor

  Production of numerous base editors and prime editors

  Base editors and prime editors are gaining more attention in gene editing era. However, their complex structures, including multiple protein and RNA components, make them challenging to express and purify.

  At GenScript, we have successfully produced numerous base editors and prime editors both in high purity and decent yield.

  

  Adenine base editor (ABE)

  

  Cytosine base editor (CBE)

  

  Prime editor (PE) 

  New! Off-the-shelf BE and PE are available here

  Type	ABE (TadA_nCas9)	CBE (APOBEC_nCas9)	PE (Cas9_RT)
  Purification procedure	Ni columna TEV protease cleavage Ni column Superdex 200 column	Ni column Q sepharose column Superdex 200 column	Ni column Superdex 200 Ni column
  Purity	>90%, by SDS-PAGE under reducing conditions.	>90%, by SDS-PAGE under reducing conditions.	>90%, by SDS-PAGE under reducing conditions.
  Yield	1.6mg/L	22.03mg/L	12.3mg/L

  

  

  
• Customized Cas12a mutant

  Customer Requirement: Deliver the Cas12a mutant and perform an activity assay

  Delivery of the high purity Cas12a mutant

  In vitro cleavage test result shows that the Cas12a mutant exhibits better cleavage activity than the wild type Cas12a

  QC report

  Concentration by Bradford	Endotoxin	Purity by SDS-PAGE	Purity by SEC-HPLC	DNase detection	Amount
  4.93mg/mL	0.648EU/mg	90%	94%	No detected	~5mg

  

  SDS-PAGESEC-HPLCDNase removal result

  In vitro cleavage test

  

  Red arrow: Full-length dsDNA substrate
  Green arrow: Fragment-1 dsDNA
  Blue arrow: Fragment-2 dsDNA

• GMP Production

  Customer Requirement:

  • Quantity: 1g, 100L fermentation of Cas variant
  • Stage 1: Plasmid and cell banking generation
  • Stage 2: Upstream and downstream process development

  Successfully delivered in 9 months

  Test result shows that the Cas variant can induced deletion of target genome in all 3 host cells efficiently.

  Test items	Specifications	Results
  Appearance	Clear, colorless liquid	PASS
  Concentration	10 ± 1 mg/mL	9.5 mg/mL
  Purity	>95%	100%
  Sterility	Negative	PASS
  Endotoxin	< 0.5 EU/mg	< 0.25 EU/mg
  Host Cell Protein	<50 ng/mg	3.9 ng/mg
  Residual DNase	< LOD	PASS
  Residual RNase	< LOD	PASS
  Host Cell DNA	<1 ng/mg	< 0.01 ng/mg

  

  Cell line1 Cell line 2 Cell line 3
• Add-on Service: In vitro Assay Development

  Cas9 nuclease in vitro cleavage test

  

  GenScript provides comprehensive solution for in vitro activity assays

  • In-house production of sgRNA and dsDNA substrate
  • Established protocols for a wide range of CRISPR systems

  Test result shows that the custom Cas9 produced by GenScript exhibited robust cleavage activity on target dsDNA

  

  Red arrow: Full-length dsDNA substrate
  Green arrow: Fragment-1 dsDNA
  Blue arrow: Fragment-2 dsDNA

Related Products and Services

Testimonials

Jack Castelli

Fred Hutchinson Cancer Center

"The people at GenScript have been tremendously helpful in providing materials for our lab. We've been able to test novel CRISPR systems such as prime editing and PASSIGE in a timely manner thanks to their work. I would recommend their services for your editing needs."

MariaMorello Miravet

Karolinska Institutet

"We are very pleased with the editing efficiency using base editors from GenScript, we got up to 100% KO!"

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