Samples were treated with DNase I (Thermo Scientific) at 37 °C for 1 h to remove any potential residual genomic DNA contamination (one unit of DNase I per 1 μg of RNA). A measure of 1 μg of total RNA was reverse-transcribed using the Superscript First Strand cDNA synthesis kit (Invitrogen) with strand-specific … See more Rosa26-CreERT2 Trf2F/F and Trf2F/+ (ref. 20) MEFs, a gift from Eros Lazzerini Denchi (The Scripps Research Institute, La Jolla, USA), were grown in DMEM supplemented with … See more Total RNA from cultured cells was extracted with RNeasy Mini Kit (Qiagen) for messenger RNA detection, or with mirVana miRNA Isolation kit (Life Technologies) for … See more Rosa26-CreERT mice (Jackson Laboratories) and Trf2 conditional knockout mice33 and mice carrying a p53 conditional allele (Jackson Laboratories) were crossed to generate Trf2/p53/Rosa26 … See more A measure of 1 μg of total cell RNA was reverse transcribed using SuperScript VILO cDNA Synthesis Kit. A volume corresponding to 10 ng of initial RNA was used for each qPCR reaction using GoTaq qPCR Master Mix … See more WebDec 1, 2024 · In both mammalian and yeast systems, telomeres are normally tethered to the nuclear envelope, where recombination is suppressed [55]. ... Taken together, DSB induction using CrispR/Cas9 does not result in limited contraction of the repeat tract but rather in expansion events or larger chromosomal deletions, but targeted use of TALENS …

Telomere identity crisis

WebSep 14, 2012 · There are two main pathways in charge of DSB repair: the non-homologous end joining (NHEJ) and the homologous recombination (HR) [ 25 ]. HR is well known for its fidelity as it uses the non-damaged sister chromatid as a template and thus is only active during the S and G2 phases. WebNov 21, 2008 · We demonstrate here that damage tolerance in stretches of long ssDNA formed at DSBs and uncapped telomeres results in extremely high frequencies of single and widely-separated multiple mutations caused by two major types of DNA damaging agents. Results DNA Damage Amplifies Hypermutability Associated with DSB Repair podcast assignment college

A model to explain how Cdc13 and Pif1 cooperate to establish the ...

WebAug 14, 2024 · CAT-R leverages the CRISPR/Cas9 system to induce a single site-specific DNA DSB using a guide RNA (gRNA) that targets the eGFP coding sequence. This design allows the mCherry coding sequence to... WebAug 27, 2024 · At the terminus of linear chromosomes, a telomere-binding protein complex, called shelterin, ensures chromosome stability by preventing chromosome … WebAug 27, 2024 · The telomere is a huge DNA-protein complex that is located at the termini of linear chromosomes. In humans, telomeric DNA comprises hexanucleotide TTAGGG … podcast assignment for middle school students