Home Resources Resources Cell Line Development 4 mainstream genome editing tools

4 mainstream genome editing tools

The mainstream genome editing tools include clustered regularly interspaced short palindromic repeat (CRISPR)/CAS9 RNA-guided nucleases, zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and helper dependent adenoviral vectors (HDAdVs). The process and protocols of mainstream genome editing methods from literatures:

1). the CRISPR-Cas9 system:

Genome engineering of mammalian haploid embryonic stem cells using the Cas9/RNA system.
The stem stable cell line development method uses the CRISPR system in combination with haploid embryonic stem cells (ESCs) to manipulate the mammalian genome.

CasOT: a genome-wide Cas9/gRNA off-target searching tool.
A local tool designed to find potential off-target sites.

Cas9 effector-mediated regulation of transcription and differentiation in human pluripotent stem cells.
A method to influence the differentiation state of human pluripotent stem cells (hPSCs) with a CRISPR-associated catalytically inactive dCas9 fused to an effector domain.

Genome-scale CRISPR-Cas9 knockout screening in human cells.
Lentiviral delivery of a genome-scale CRISPR-Cas9 knockout (GeCKO) library targeting 18,080 genes with 64,751 unique guide sequences enables both negative and positive selection screening in human cells.

Genetic screens in human cells using the CRISPR-Cas9 system.
A pooled, loss-of-function genetic screening approach suitable for both positive and negative selection that uses a genome-scale lentiviral single-guide RNA (sgRNA) library containing 73,000 sgRNAs to generate knockout collections and performed screens in two human cell lines.

Genome engineering using the CRISPR-Cas9 system.
A set of tools for Cas9-mediated genome editing via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, as well as generation of modified cell lines for downstream functional studies. It takes 1-2 weeks to do target design and gene modifications, and 2-3 weeks to obtain modified clonal cell lines.

CRISPR Genome Engineering Resources
Comprehensive CRISPR genome engineering resources organized by Zhang’s lab.

Cas9 as a versatile tool for engineering biology.
Advances of the Cas9 targeting methodology, and potential applications ranging from basic science to the clinic.

2). zinc finger nucleases (ZFNs):

Gene editing in human stem cells using zinc finger nucleases and integrase-defective lentiviral vector delivery.

3). transcription activator-like effector nucleases (TALENs):

Genetic engineering of human pluripotent cells using TALE nucleases.

4). helper dependent adenoviral vectors (HDAdVs):

 

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We index the protocols for your information only. It does not necessarily mean that we agree with all of them. You rather than Syd Labs takes full responsibility for using any information described here.