Recent advancements in targeted gene editing and genome editing techniques have allowed scientists to choose from multiple molecular approaches for highly-specific, targeted DNA modifications to their gene(s) of interest in cell lines or mouse models. These techniques, including leveraging CRISPR/Cas9 nuclease, transcription activator-like effector-based nucleases (TALEN), and zinc finger nucleases (ZFNs), have allowed scientists to undertake genome editing experiments with unparalleled ease and specificity, ushering in a new era of widespread availability of genome engineering techniques and applications.
cell line engineering and Genome Editing
Genome editing and gene editing techniques are increasingly being used in gene knockout and gene mutagenesis studies for engineering cell lines and mouse models. The CRISPR/Cas9 system in particular has been widely adopted to undertake gene knockout experiments in which a true knockout effect can be achieved in stable cell lines, as compared to traditional, transient and incomplete knockdown techniques such as RNAi. CRISPR technology is frequently leveraged for reversible gene knockdown, through CRISPR interference (CRISPRi) and gain-of-function gene knockin approaches are made possible by the CRISPR/Cas9 system as well. The precision and relative simplicity of the CRISPR gene editing system has enabled scientists to modify DNA quickly and efficiently in both engineered cell lines and mouse models, with a wide range of possible applications across research areas, target genomes, and industries. Gene editing has the potential to revolutionize genetic approaches to personalized medicine, and could enable a new era of gene-based therapeutic treatment options for heretofore untreatable diseases.
CRISPR Workflow and GENEWIZ Services
GENEWIZ is pleased to provide a number of services to expedite research in genome editing. The diagram below illustrates a typical CRISPR workflow and the steps within this workflow where GENEWIZ can help move your experiment forward.
Current examples of genome editing and gene editing applications in research include:
Cancer researchers investigate a candidate gene’s involvement in cancer progression using the CRISPR gene editing system to eliminate gene functionality and study the downstream effects in edited, stable cell lines.
Malaria and other mosquito-borne disease researchers leverage the CRISPR genome editing system to disrupt genes that control female mosquito reproduction, resulting in the possibility of a genetic approach to mosquito population control for regions susceptible to outbreaks of mosquito-borne diseases.
Stem cell researchers leverage CRISPR interference (CRISPRi) to reversibly knockdown expression of target genes in induced pluripotent stem cells (iPSCs) to study the effect on subsequent developmental pathways.
Biofuel crop scientists employ TALE-nucleases (TALENs) to disrupt highly redundant or polyploid lignin genes in the sugarcane genome to increase availability of cell wall sugars for enhanced production of ethanol in biofuel generation.
Outsource your CRISPR construct (both gRNA and donor construct) synthesis projects to the Gene Synthesis industry leader in quality and customer service. A variety of different synthesis options for CRISPR constructs are available, including:
CRISPR gRNA Construct Synthesis Service
Your design, our guarantee. GENEWIZ’s guide RNA (gRNA) construct synthesis service provides gRNA constructs cloned into your custom vector of choice with a quick turnaround time and low flat price per construct. You provide the sequence, GENEWIZ provides the custom synthesis, cloning, and sequence verification for one low price. Need a custom vector synthesized? Take advantage of GENEWIZ’s expertise, quality, and reliability with difficult constructs for your custom vector synthesis projects as well. Learn more >
Looking to create a gRNA library to screen knockouts / knockins of a variety of genes in one experiment? GENEWIZ custom synthetic library service can create a custom gRNA library for high-throughput CRISPR screening needs. Learn more >
CRISPR and Genome Editing Validation Applications
GENEWIZ provides a variety of on-target and off-target validation approaches, leveraging both Sanger Sequencing and Next Generation Sequencing.
Next Generation Sequencing
Upon completion of genome editing experiments, scientists need to evaluate and confirm on- and off-target effects before proceeding with any downstream applications. Genome editing efficiency can be determined for pools of transfectants or assessed after cloning. Whether you are interested in confirming on-target / off-target effects via amplicon sequencing, transcriptome effects via RNA-seq, or genome-level effects via whole genome sequencing, the GENEWIZ NGS team has solutions to analyze the success of your CRISPR experiments. RNA-Seq > Amplicon Sequencing > Whole Genome Sequencing >
Sanger confirmation of on-target genome editing effects is simple, fast, and easy. Let the Sanger expert provide rapid turnaround and reliable results for Sanger validation of CRISPR constructs for experiments big and small. COMING SOON – GENEWIZ U6 Universal Primer! Learn more >
PCR + Sequencing CRISPR Analysis Package
Let the Sanger experts amplify, purify, and sequence your specified on-target or off-target regions of interest from your CRISPR experiment. GENEWIZ's SNP / Mutation Analysis service includes PCR validation, amplicon production and purification, as well as Sanger sequencing for your CRISPR regions of interest. Simply identify your region of interest, and GENEWIZ will take it from there. Nucleic acid extraction can also be added to any SNP / Mutation Analysis project. Learn more >