ALK Gene Editing

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ALK Gene Editing    

ALK (Anaplastic lymphoma kinase) is also named CD246, and belongs to the tyrosine kinase family. The ALK gene is located on the human chromosome at 2p23.2-p23.1, and normally expressed in the brain, small intestine, and testis, but not in the normal lymphoid cells. A recent study shows that longer heparin chains can be used as ligands for ALK to induce ALK dimerization and self-phosphorylation, which stimulates downstream signaling by activating PI3K/AKT, MAPK/ERK, and STAT3 pathways, leading to cancer cell growth, survival and metastasis (Figure 1).

 ALK signaling in human/mouseFigure 1: ALK signaling in human/mouse

ALK plays an important role in the development and function of the nervous system, controls the basic mechanisms of cell proliferation, survival and differentiation in response to extracellular stimuli. ALK can be oncogenically altered in several malignancies including neuroblastoma, non-small cell lung cancer (NSCLC) and anaplastic large cell lymphoma (ALCL). The most common ALK change is chromosomal rearrangement, resulting in fusion genes, as seen in ALCL and NSCLC (Table 1). In other tumors, the increasing of ALK gene copy number and ALK activating mutation have also been reported. Clinical trials have demonstrated that ALK is effective as a new target for tumor therapeutics. However, drug resistance is universal. A variety of mechanisms about the resistance of ALK to TKI have been discovered, which are providing information for the development of new therapeutic strategies to overcome clinical drug resistance and useful to develop newer ALK inhibitors and other targeting strategies.

Table 1: ALK chromosomal translocations in tumors (Vijaykumar R. Holla.)

Tumor typesFusion
ALCLATIC-ALK; CLTC-ALK; NPM1-ALK; TFG-ALK; TPM3-ALK; TPM4-ALK
IMTCLTC-ALK; DCTN1-ALK; TPM3-ALK; RANBP2-ALK; TPM4-ALK
LungKIF5B-ALK; KLC1-ALK
NSCLCEML4-ALK
CRCCAD-ALK
ThyroidSTRN-ALK
BCLSQSTM1-ALK; CLTC-ALK
OvarianFN1-ALK

ALK Gene Editing Service

CRISPR/Cas9 PlatformCB, one of the leading genomic editing companies, has focused on building efficient systems and procedures to meet the needs and timelines of clients working in the CRISPR/Cas9 gene editing. Our scientists have deep gene editing knowledge and extensive experience in experimental operation and data processing. Based on our platform, CRISPR/Cas9 PlatformCB offers the CRISPR/Cas gene editing services in cell lines and animal models, which allow you achieve specific gene knockout, knockin, or point mutation in vitro and in vivo within a shorter timescale.

  • ALK Gene Editing Cell Line Generation

Our scientists are very familiar with the culture methods and genetic backgrounds of various cell lines and have successfully implemented gene editing of ALK in both easy-to-transfect cell lines and hard-to-transfect cells. Tell us the cell lines your project needs, we will offer you a one-stop-shop ALK gene editing service from strategy design to final stable cell lines. And our ALK gene editing cell line generation services include:

✧ SgRNA design and synthesis
✧ Transfect the cell line you interest
✧ Select the high expression cell and sort monoclonal cell
✧ Validate the knockout/knockin/point mutation of ALK by PCR and sequencing
✧ Produce cryogenic preserved vials of stable cells and a final report

Typically, we develop CRISPR-mediated gene editing cell lines including HEK239T, Hela, HepG2, U87, but we can use other cell lines according to your requirements.

Host cell line: Ba/F3, CHO, MDA-MB-453, MDA-MB-231NIH3T3, T47D, Neuro2a, MCF7, RKO, K562, RAW264.7, etc.

  • ALK Gene Editing Animal Model Generation

CRISPR/Cas9 PlatformCB also has extensive experience in incorporating CRISPR-Cas9 technology into animal models, which have been recognized by our clients. We provide you with CRISPR/Cas ALK gene editing animal models at a reasonable cost and in a shorter time and guarantee at least 2 founders or 3 F1 animals. Our ALK gene editing animal model generation services include:

➢ ALK gene conventional knockout animals
➢ ALK gene conditional knockout animals
➢ ALK point mutation animals
➢ ALK knockin animals

Alternative species: mouse, rat, rabbit, zebrafish, C. elegans, etc.

Tell us your needs, we will provide you a custom one-stop-shop CRISPR/Cas9 gene editing services containing strategy design, gRNA, and Ca9 mRNA construction, transfection or injection and final cell lines or F1 animals. We guarantee our clients the most reliable and efficient research services to best match your research goals, and faster turnaround times, lower prices.

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References:

  1. Morris SW. et al. Fusion of a kinase gene, ALK, to a nucleolar protein gene, NPM, in non-Hodgkin's lymphoma. Science. 1994; 263: 1281–1284.
  2. Murray PB. et al. Heparin is an activating ligand of the orphan receptor tyrosine kinase ALK. Sci Signal. 2015; 8(360): ra6.
  3. Yao S. et al. Anaplastic lymphoma kinase is required for neurogenesis in the developing central nervous system of zebrafish. PLoS One. 2013; 8(5): e63757.
  4. Vijaykumar R. Holla. et al. ALK: a tyrosine kinase target for cancer therapy. Cold Spring Harb Mol Case Stud. 2017; 3(1): a001115.
  5. Jessica J. Lin, et al. Targeting ALK: Precision Medicine Takes on Drug Resistance. Cancer Discov. 2017 Feb; 7(2): 137–155.  
For research use only. Not intended for any clinical use.

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