Generation and Characterization of a Transgenic Mouse Carrying a Functional Human β-Globin Gene with the IVSI-6 Thalassemia Mutation

Table 5

Strengths of the normal and cryptic splicing sites generated by the β⁺IVSI-6 thalassemic mutation.

Splicing site	Sequence	Base pairs	Strength			Amplicon size	Comments/Hypotheses
			(a)	(b)	(c)
+1; (normal)		7+; 3+	86.64	8.08	0.64	153 bp
+1; (mutated; IVSI-6)		6+; 2+	84.46	5.52	0.14	153 bp
−38; (cryptic)		5+; 1	83.50	5.54	0.21	115 bp	PTC. Unstable transcript, low sensitivity to NMD; (in comparison with the −16 cryptic splicing site)
−16; (cryptic)		5+; 2	90.40	6.13	0.54	137 bp	PTC. Unstable transcript, sensitive to NMD
+13; (cryptic)		5+; 1+	79.67	−0.83	0.46	165 bp	No PTC. GT located 7 nucleotides downstream the IVSI-6 mutation

Sequences of the normal and cryptic splicing sites generated by the β⁺IVSI-6 thalassemic mutation. The cryptic donor GU (boxed) sites are numbered with respect to the +1 position of the normal one. Potential Watson-Crick base pairs to U1 (upper) and U6 (lower) are quantified: (+) indicates a G/U wobble base pair. Strengths of donor splicing sites are expressed as scores calculated with Human Splicing Finder Matrices from http://www.umd.be/HSF/ (a) or MaxEntScan from http://www.umd.be/HSF/ (b) or from http://www.fruitfly.org/seq_tools/splice.html (c).