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With thousands of patients worldwide, CAPN3 c.550delA is the most frequent mutation causing severe, progressive, and untreatable limb girdle muscular dystrophy. We aimed to genetically correct this founder mutation in primary human muscle stem cells. We designed editing strategies providing CRISPR-Cas9 as plasmid and mRNA first in patient-derived induced pluripotent stem cells and applied this strategy then in primary human muscle stem cells from patients. Mutation-specific targeting yielded highly efficient and precise correction of CAPN3 c.550delA to wild-type for both cell types. Most likely a single cut generated by SpCas9 resulted in a 5’ staggered overhang of one base pair, which triggered an overhang-dependent base replication of an A:T at the mutation site. This recovered the open-reading frame and the CAPN3 DNA sequence was repaired template-free to wild-type, which led to CAPN3 mRNA and protein expression. Off-target analysis using amplicon sequencing of 43 in silico predicted sites demonstrates safety of this approach. Our study extends previous usage of single cut DNA modification since our gene product has been repaired into the wild-type CAPN3 sequence with the perspective of a real cure.

"Una modernidad política iberoamericana. Siglo XIX. Formación, relaciones internacionales y representaciones de la nación" is an edited volume by the historians Pierre Géal, Sebastián Martínez, Graziano Palamara and Daniel Rojas. The text is part of the Contemporary History of America collection of Editorial Marcial Pons and the Institute of Historical Research of the Universidad Michoacana de San Nicolás de Hidalgo. The ai...

En La Paz, Baja California Sur, en el ciclo 1991/1992 se evaluaron tres grupos de líneas de triticale (X Triticosecale Wittmack) y uno de centeno (Secale cereale), con el objeto de seleccionar líneas con potencial forrajero y de grano que se adapten a las condiciones ambientales de esta zona árida e identificar nuevas fuentes de variabilidad genética para el programa de investigación local. El diseño experimental fue bloques completos al azar con tres repeticiones. Se incluyeron 160, 45, 205 y 46 líneas de triticales de primavera, triticales de invierno, triticales facultativos y centenos, respectivamente. Del grupo de triticale de primavera se seleccionaron 22 líneas para forraje y 53 líneas para grano, con rendimientos superiores a 31 y 4.0 t ha-1 de forraje y grano, respectivamente. De los triticales de invierno se seleccionaron 34 líneas para forraje y 4 líneas para grano con rendimientos superiores a 31 y 3.0 t ha-1 de forraje y grano. De los triticales facultativos se seleccionaron 83 líneas para forraje y 18 líneas para grano con rendimientos superiores a 31 y 4.0 t ha-1 de forraje y grano. De los centenos se seleccionaron 13 líneas para forraje y 3 líneas para grano con rendimientos superiores a 31 y 1.5 t ha-1 de forraje y grano.

Skeletal muscle can regenerate from muscle stem cells and their myogenic precursor cell progeny, myoblasts. However, precise gene editing in human muscle stem cells for autologous cell replacement therapies of untreatable genetic muscle diseases has not yet been reported. Loss-of-function mutations in SGCA, encoding α-sarcoglycan, cause limb-girdle muscular dystrophy 2D/R3, an early onset, severe and rapidly progressive form of muscular dystrophy affecting equally girls and boys. Patients suffer from muscle degeneration and atrophy affecting the limbs, respiratory muscles, and the heart. We isolated human muscle stem cells from two donors with the common SGCA c.157G>A mutation affecting the last coding nucleotide of exon 2. We found that c.157G>A is an exonic splicing mutation that induces skipping of two co-regulated exons. Using adenine base editing, we corrected the mutation in the cells from both donors with >90% efficiency, thereby rescuing the splicing defect and α-sarcoglycan expression. Base edited patient cells regenerated muscle and contributed to the Pax7 positive satellite cell compartment in vivo in mouse xenografts. We hereby provide the first evidence that autologous gene repaired human muscle stem cells can be harnessed for cell replacement therapies of muscular dystrophies. ONE SENTENCE SUMMARY: Patient primary muscle stems cells gene repaired with >90% efficiency by base editing maintain their regenerative properties for autologous cell replacement therapies of muscular dystrophy.

El estrés es un tema de interés hoy en día debido a su alta persistencia y su relación con enfermedades crónicas y otros hallazgos que la ciencia actual ha permitido saber. Con el interés de conocer la presencia de estrés a través de sus manifestaciones, se ha diseñado un sistema electrónico e informático que mide las respuestas fisiológicas al estrés: frecuencia cardíaca (FC, frecuencia cardíaca), presión arterial (PA, presión arterial) y actividad electrodérmica (EDA) ) Esta es una investigación explicativa de un tipo experimental con dos niveles de variable independiente (presencia de estresor y ausencia de estresor), que consta de ocho participantes. Los resultados de las diferencias identificadas entre el control y los grupos experimentales, especialmente en la medición de la presión arterial sistólica y diastólica; y dentro del grupo experimental difieren en el EDA entre el momento anterior y la exposición al factor estresante, así como en la PA sistólica entre el momento de exposición al factor estresante y el posterior. Stress is a topic of interest today due to its high persistence and its relationship with chronic diseases and other findings that current science has allowed to know. With the interest of knowing the presence of stress through its manifestations, an electronic and computer system that measures physiological responses to stress has been designed: heart rate (HR, heart rate), blood pressure (BP, blood pressure) and Electrodermal Activity (EDA). This is an explanatory investigation of an experimental type with two levels of independent variable (presence of stressor and absence of stressor), consisting of eight participants. The results of the differences identified between the control and experimental groups, especially in the measurement of systolic and diastolic BP; and within the experimental group they differ in the EDA between the previous moment and the exposure to the stressor, as well, as in the systolic BP between the moment of exposure to the stressor and the subsequent one.

Additional file 5. pathway_components. Abstract of the WNT and BCR signaling pathway and the components.

Additional file 3. supplement figures. Heatmaps of the top-100 up- and downregulated genes of the biological triplicates.