Ligase chain reaction assay for human mutations: the Sickle Cell by LCR assay

Abstract We can detect the β-globin gene sickle cell mutation by using an assay based on the ligase chain reaction. The simultaneous amplification of the human growth hormone gene in the same reaction serves as a control for the amount of template DNA or amplification efficiency. Ligation products, which are biotinylated at one end and tagged with an arbitrary “tail” sequence at the other, are captured by hybridization to “tail”-complementary oligonucleotides immobilized on polystyrene microwells. The captured ligation products are detected colorimetrically by use of streptavidin–alkaline phosphatase conjugate. In a study of 24 subjects, the assay unequivocally discriminated among normal, carrier, and sickle cell genotypes.

Download Full-text

Reliability of detection by polymerase chain reaction of the sickle cell-containing region of the β-globin gene in single human blastomeres

Human Reproduction ◽

10.1093/oxfordjournals.humrep.a137710 ◽

1992 ◽

Vol 7 (5) ◽

pp. 630-636 ◽

Cited By ~ 11

Author(s):

Susan J. Pickering ◽

Josie M. McConnell ◽

Martin H. Johnson ◽

Peter R. Braude

Keyword(s):

Polymerase Chain Reaction ◽

Sickle Cell ◽

Globin Gene ◽

Chain Reaction ◽

Polymerase Chain

Download Full-text

Improved diagnosis of sickle cell mutation by a robust amplification refractory polymerase chain reaction

Clinical Biochemistry ◽

10.1016/s0009-9120(99)00071-5 ◽

1999 ◽

Vol 32 (8) ◽

pp. 677-680 ◽

Cited By ~ 2

Author(s):

Mehmet Simsek ◽

Shahina Daar ◽

Hana Ojeli ◽

Riad Bayoumi

Keyword(s):

Polymerase Chain Reaction ◽

Sickle Cell ◽

Chain Reaction ◽

Cell Mutation ◽

Polymerase Chain

Download Full-text

Correction of the Sickle Cell Mutation Through Base and Prime Editing in Hematopoietic Stem Cells

10.20944/preprints202009.0490.v1 ◽

2020 ◽

Author(s):

Kelly Ji

Keyword(s):

Sickle Cell Disease ◽

Sickle Cell ◽

Globin Gene ◽

Treatment Options ◽

Point Mutations ◽

Inherited Disease ◽

Cell Disease ◽

Hematopoietic Stem ◽

Base Editing ◽

Cell Mutation

Sickle cell disease is characterized by stiff, “sickled” red blood cells that have difficulty moving through the bloodstream and do not efficiently carry oxygen. It is an inherited disease with severely limited treatment options, and is caused by a point mutation. Its prevalence in black and brown communities makes the already limited treatment options even less accessible. Base editing and prime editing are two relatively recent discoveries in the field of genome editing and were developed after the groundbreaking discovery of the CRISPR Cas9 system. While not fully tested, they hold a lot of promise in providing alternative treatment options for sickle cell disease. Both editing systems are able to install individual point mutations in the beta globin gene, which is where the sickle cell mutation occurs, and can thus cure sickle cell disease (in theory). In this paper we outline the mechanisms of CRISPR-Cas9 systems and base and prime editing, and provide insight into how to apply them to treat SCD. Further investigation should be done on specific editing systems and designs to use to ensure optimal treatment of SCD.

Download Full-text