Gene Editing Solutions

Introduction

Traditional CRISPR editing techniques involve transfecting expression plasmids encoding guide RNA and/or Cas9 nuclease into host cells. By combining Cas9 protein with synthesized synthetic single guide RNAs (sgRNAs) into ribonucleoprotein (RNP) prior to delivery can significantly simplify the experimental process, while improving editing efficiency, reducing off-target effects, and avoiding immune response.

GenScript offers sgRNA synthesis service with guaranteed sequence correctness. GenCRISPR synthetic sgRNAs are more efficient, less toxic and safer than plasmid or virus based editing solutions, and are proven effective for targeted gene editing in knockout and HDR mediated knock-in experiments. We also provide custom synthesis of pegRNA for prime editing and crRNA for Cpf1/Cas12a editing systems.

sgRNA Synthesis

• GenCRISPR Synthetic sgRNA
• GenTitan 8 Million Oligo Pool
• Custom synthesis of pegRNA / crRNA
• cGMP guide RNA

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Gene Editing Efficiency Assessment

• GenCrispr sgRNA Screening Kit
• GenCRISPR™ Mutation Detection Kit
• GenCRISPR™ Antibodies
• NGS Target Enrichment solutions

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GenCRISPR Synthetic sgRNA

Synthetic single guide RNA (sgRNA) is sgRNA that is chemically synthesized using the latest technological advancements. Utilizing synthetic sgRNA is a simple and cost-effective way to improve your CRISPR editing, as it offers higher editing efficiency, lower cytotoxicity, and higher lot-to-lot stability compared to IVT sgRNA.

Additionally, we offer custom synthesis of pegRNA for prime editing and crRNA for Cpf1/Cas12a editing systems.

Introduction

Through our partnership with Feng Zhang's laboratory at the Broad Institute of MIT and Harvard*, GenScript offers validated GMP Grade and RUO Grade Cas9, Cas12a and Cas13a nuclease products, sgRNA synthesis and HDR donor template synthesis services, and resources to help with leveraging CRISPR gene editing for basic research and clinical manufacturing.

One powerful application of CRISPR/Cas genome editing technology involves the precise insertion of DNA sequences via the homology-directed repair (HDR) pathway, and choosing the right HDR template is crucial for the efficacy of this approach. GenScript offers high quality, sequence verified HDR templates in both linear and circle format for maximizing the editing efficiency of your CRISPR experiments.

CRISPR/Cas9 technology was used to knock-out genes and knock-in the target sequence, resulting in gene editing cell lines that can be stably passaged. GenScript provides a variety of CRISPR gene editing cell lines. Utilizing the advantages of chemically synthesized long single-stranded gRNA, our RNP delivery system achieves both high editing efficiency and low toxicity, and support multiple delivery systems of lentivirus and plasmid to provide gene-editing cell line services with highly customizable target genes, editing regions and cell types.

Cas Enzymes

• Cas Nuclease Product List
• Cas9
• Cas12a
• Prime editors
• Base editors
• Integrated CRISPR/Cas Protein Solution
• Catalog IVT mRNA Products

HDR Templates

• Gene Synthesis Service
• CRISPR HDR knock-in templates (ssDNA, dsDNA, circle dsDNA)
• cGMP HDR template

Gene Editing Modeling Services

• Customized Stable CRISPR Cell Line Development
• Microbial Genome Editing Service
• CAR-T knock in optimization kits

GenCRISPR™ Cas Proteins

GenCRISPR™ Cas9 proteins are optimized for high CRISPR editing efficiency across various gene targets and cell types.

GenScript offers a range of Cas9 proteins, including GenCRISPR™ SpCas9, GenCRISPR™ Ultra eSpCas9, and GMP-grade Cas9.

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GenCRISPR™ SpCas9

Wild-type Cas9 protein engineered for optimal CRISPR editing performance

Applications: Suitable for a wide range of CRISPR gene editing experiments.

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GenCRISPR™ Ultra eSpCas9

High-fidelity Cas9 variant engineered for enhanced specificity without compromising efficiency

Applications: Ideal for CRISPR experiments that need precise editing or low off-target effects.

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GMP-grade GenCRISPR™ SpCas9 and Ultra eSpCas9

Wild-type and high-fidelity Cas9 produced to meet the requirements for clinical development

Applications: Ideal for CRISPR projects that aim for therapeutic applications.

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CRISPR Cas Nuclease Products

Through our partnership with Feng Zhang's laboratory at the Broad Institute of MIT and Harvard*, GenScript offers validated GMP Grade and RUO Grade Cas9, Cas12a and Cas13a nuclease products, sgRNA synthesis and HDR donor template synthesis services, and resources to help with leveraging CRISPR gene editing for basic research and clinical manufacturing.

Catalog IVT mRNA, circRNA & saRNA Products

• Catalog CAS enzymes mRNA

• IVT mRNA Synthesis

In vitro transcription (IVT) refers to the process by which the DNA template containing the protein sequence is transcribed into mRNA using RNA polymerase. When this mRNA is delivered into host cells, it serves as the blueprint for protein synthesis.

GenScript provides off-the-shelf IVT CRISPR Cas mRNA products in linear mRNA and circular RNA formats for various applications.

Name	Cat. No	Description
eSpCas9 Circular RNA (CVB3)	RP-A00017	The eSpCas9 mRNA expresses enhanced-specificity SpCas9 protein.	Learn More
eSpCas9 mRNA (Cap1, m1Ψ)	RP-A00018	The eSpCas9 mRNA expresses enhanced-specificity SpCas9 protein. It has 100% substituted with N1-methyl-pseudo Uridine for enhanced expression and reduced immunogenicity.
GenLNP-A01-eSpCas9 mRNA (m1Ψ)-TRAC sgRNA	RP-A00019	The ALC0315 LNP formulation loaded with eSpCas9 mRNA (m1Ψ) and TRAC sgRNA.
GenLNP-S01-eSpCas9 mRNA (m1Ψ)-TRAC sgRNA	RP-A00020	The SM102 LNP formulation loaded with eSpCas9 mRNA (m1Ψ) and TRAC sgRNA.
eSpCas9 mRNA (Cap1, 5-MOU)	RP-A00050	The eSpCas9 mRNA expresses enhanced-specificity SpCas9 protein. It has 100% substituted with 5-methoxyuridine forenhanced expression and reduced immunogenicity.
CPF1 mRNA (N1-Methylpseudouridine/m1Ψ)	RP-A00004	Cpf1 is a novel class of CRISPR-Cas DNA endonuclease. The LbCpf1/Cas12a allow efficient mutagenesis in zebrafish and Xenopus (Moreno-Mateos, et al. Nat Communication, 2017; 8: 2024). LbCpf1 increases homology-directed genome editing.
GenScript PE2/PE3 mRNA (N1-Methylpseudouridine/m1Ψ)	RP-A00044	Prime editing (PE) systems minimally consist of two components: a programmable DNA nickase fused to an engineered reverse transcriptase and a pegRNA. The PE2/PE3 mRNA sequence was from publication: Nelson, et al. Nat Biotechnology, 2022; 40: 402-410.

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		Linear mRNA		Self-Amplifying RNA		Circular RNA
	Structure	Linear; Open 5’ and 3’ end mRNA exonuclease degradation		Linear; Open 5’ and 3’ end mRNA exonuclease degradation		Close loop; No free 5’ 3’ end Eliminating mRNA exonuclease degradation
	Ribosome recruiting	5’ Cap: Initiates translation & stabilizes mRNA		5’ Cap: Initiates translation & stabilizes mRNA		Internal Ribosome Entry Site (IRES) : Initiates translation
	ORF	Codon optimization for translation yield Modified nucleosides, eg. M1Ψ Insert < 20 kb		Codon optimization for translation yield Modified nucleosides, eg. M5C Insert < 6 kb		Codon optimization for translation yield Insert < 4 kb
	UTR	Regulatory elements in the 5’ and 3’-UTR stabilize mRNA and increase translation initiation		Replicon for further self-replication of the mRNA		Scarless circRNA options for coding and non-coding applications
	PolyA tail	Stabilizes RNA molecule & Prevents degradation		Stabilizes RNA molecule & Prevents degradation		Not required
	Protein Expression	+++ Strong		+++++ Strongest		++++ Stronger
	Expression half-life	+++ Shorter		+++++ Longest		++++ Longer
	Immunogenicity	++++ Low		++ High		+++ Medium
	Dosage requirement	++ High		+++++ Lowest		+++ Medium
	Novelty	+++ Traditional		++++ Novel		+++++ Novel

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Homology-Directed Repair (HDR) Knock-in Templates

GenExact™ ssDNA
Precise KI and low cytotoxicity

Application

Ideal for T cell therapy

Ideal for cell/animal model generation

Advantages

Minimal cytotoxicity

Precise knock-in, minimized off-target effects

High purity and sequence-verified

Specifications

Insertion length: 200*-6,000 nt

µg to mg scale

Research to cGMP grade

*We also offer <200 nt ssDNA through our DNA Oligo Synthesis Service.

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GenWand™ dsDNA
Long gene knock-in in large scale

Application

Ideal for long gene CRISPR KI

Ideal for screening and scale-up

Advantages

Covalently closed ends protect for better accuracy

More suitable for scale-up

High purity and sequence-verified

Specifications

Insertion length: 1-10 kb

µg to g scale

Research to cGMP grade

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GenCircle™ dsDNA
Circular template with no resistance gene

Application

Ideal replacement for plasmid HDR template

Ideal for scale-up and fast delivery

Advantages

KI efficiency increase by up to 30%

429bp backbone, lower cytotoxicity & higher transfection efficiency

No antibiotic resistance avoids regulatory concern

High purity and sequence-verified

Specifications

Insertion length: 1-20 kb

µg to g scale

Research to cGMP grade

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GenScript provides over 20,000 pLentiCRISPR v2 plasmids containing gRNA sequences, and all sgRNA sequences have been verified by MIT. You can find the relevant gRNA by searching for Gene Name, Gene Symbol, or Gene ID in the database. We also provide empty vector service with vector sequences that have been verified by MIT. Multiple vector types including binary vector and all-in-one are available, and resistance screening methods can be selected or modified according to your needs.

GenScript now offers lipid nanoparticle (LNP) formulation services for the delivery of mRNA and circular RNA. Our Ready-Edit CRISPR mRNA-LNP Solutions combines popular CRISPR Cas mRNAs with different LNP formulation as off-the-shelf products to offer highly efficient, precise, and safe editing experience. Utilizing state-of-the-art lipid formulations and targeting ligands, our Targeted LNP Service ensures that the payloads are delivered directly to specific cells or tissues, thereby maximizing therapeutic impact and minimizing off-target effects.

AAV (Adeno-Associated Virus) and LVV (Lentiviral Vector) are two important viral vectors used in gene editing and therapy development. GenScript provides services covering from upstream viral vector plasmid construction to downstream viral vector packaging for research, preclinical study and clinical development.

Plasmid Services

• Plasmid DNA Preparation
• Plasmid CDMO Services
• GenCRISPR™ gRNA/Cas9 Plasmids
• mRNA plasmids
• AAV plasmids
• Cell free mRNA template

Non-Viral Delivery (LNPs)

• IVT mRNA synthesis
• GMP-like mRNA synthesis
• Ready-Edit LNP Solutions
• Lipid Nanoparticle Formulation for Advanced RNA Delivery
• Targeted LNP Formulation
• Catalog LNP Products

Viral Delivery

• AAV Packaging Service (Research Grade)
• AAV CDMO Services
• Lentivirus Packaging Service (Research Grade)
• Lentiviral Vector CDMO Services

• ReadyEdit LNP Solutions Offerings
  

  1. Start your pilot project with ReadyEdit LNP Solutions

  • Cas9 system
  • Cas12 system
  • Base and Prime Editing.

  

  2. Build your CRISPR components including the synthesis of mRNA, sgRNA, ssDNA, and more!

  

  3. LNP Formulations validated and optimized for specific CRISPR applications to significantly enhance your project's success rate.

  

  4. Validation with robust screening platforms to validate your gene-editing LNP designs.

  Key Benefits and Advantages of Ready-Edit LNP Platform

  

  Targeted Delivery

  LNPs can be engineered to enhance the delivery efficiency of CRISPR system payloads

  

  Reduced Off-Target Effects

  mRNA provides transient expression of the gene-editing tool, reducing the likelihood of off-target effects

  

  Scalability and Production

  synthesized quickly and in large quantities using in vitro transcription

  

  Safety Profile

  mRNA does not integrate into the host genome, reducing the risk of insertional mutagenesis

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Viral Packaging Workflow