Maximizing the Efficacy of CRISPR/Cas Homology-Directed Repair Gene Targeting

Clustered regularly interspaced short palindromic repeats/CRISPR-associated system (CRISPR/Cas) is a powerful gene editing tool that can introduce double-strand breaks (DSBs) at precise target sites in genomic DNA. In mammalian cells, the CRISPR/Cas-generated DSBs can be repaired by either template-free error-prone end joining (e.g., non-homologous end joining/microhomology-mediated end joining [NHEJ]/[MMEJ]) or templated error-free homology-directed repair (HDR) pathways. CRISPR/Cas with NHEJ/MMEJ DNA repair results in various length insertions/deletion mutations (indels), which can cause frameshift mutations leading to a stop codon and subsequent gene-specific knockout (i.e., loss of function). In contrast, CRISPR/Cas with HDR DNA repair, utilizing an exogenous repair template harboring specific nucleotide (nt) changes, can be employed to intentionally edit out or introduce mutations or insertions at specific genomic sites (i.e., targeted gene knock-in). This review provides an overview of HDR-based gene-targeting strategies to facilitate the knock-in process, including improving gRNA cleavage efficiency, optimizing HDR efficacy, decreasing off-target effects, suppressing NHEJ/MMEJ activity, and thus expediting the screening of CRISPR/Cas-edited clonal cells

El mapa perceptual del consumidor de microcréditos para la ciudad de Bogotá

Microcredit typically refers to very small loans for unsalaried borrowers with little or no collateral and has been seen as a financing option to encourage the productive sector and support the brain capital of the population who has no typical bank loans. By mapping consumer microcredit perceptual in Bogota we could infer three dimensions projected: working capital, permanence and trust-worthiness, contrasting those people who do not have credit or any bank loans but take strongly into account just a dimension.El microcrédito se presenta como una opción de financiamiento para estimular el sector productivo y apoyar el capital intelectual de la población que no tiene oportunidad de acceso a los créditos bancarios convencionales. A partir de la elaboración del mapa perceptual del consumidor de microcréditos de la ciudad de Bogotá, se llegó a la conclusión de que este valora tres dimensiones: capital de trabajo, permanecía y confianza, mientras que los individuos que no han adquirido crédito o que han empleado créditos bancarios solo valoran una dimensión fuertemente.

Intronic Polyadenylation in Acquired Cancer Drug Resistance Circumvented by Utilizing CRISPR/Cas9 with Homology-Directed Repair: The Tale of Human DNA Topoisomerase IIα

Intronic polyadenylation (IPA) plays a critical role in malignant transformation, development, progression, and cancer chemoresistance by contributing to transcriptome/proteome alterations. DNA topoisomerase IIα (170 kDa, TOP2α/170) is an established clinical target for anticancer agents whose efficacy is compromised by drug resistance often associated with a reduction of nuclear TOP2α/170 levels. In leukemia cell lines with acquired resistance to TOP2α-targeted drugs and reduced TOP2α/170 expression, variant TOP2α mRNA transcripts have been reported due to IPA that resulted in the translation of C-terminal truncated isoforms with altered nuclear-cytoplasmic distribution or heterodimerization with wild-type TOP2α/170. This review provides an overview of the various mechanisms regulating pre-mRNA processing and alternative polyadenylation, as well as the utilization of CRISPR/Cas9 specific gene editing through homology directed repair (HDR) to decrease IPA when splice sites are intrinsically weak or potentially mutated. The specific case of TOP2α exon 19/intron 19 splice site editing is discussed in etoposide-resistant human leukemia K562 cells as a tractable strategy to circumvent acquired TOP2α-mediated drug resistance. This example supports the importance of aberrant IPA in acquired drug resistance to TOP2α-targeted drugs. In addition, these results demonstrate the therapeutic potential of CRISPR/Cas9/HDR to impact drug resistance associated with aberrant splicing/polyadenylation

El mapa perceptual del consumidor de microcréditos para la ciudad de Bogotá

Microcredit typically refers to very small loans for unsalaried borrowers with little or no collateral and has been seen as a financing option to encourage the productive sector and support the brain capital of the population who has no typical bank loans. By mapping consumer microcredit perceptual in Bogota we could infer three dimensions projected: working capital, permanence and trust-worthiness, contrasting those people who do not have credit or any bank loans but take strongly into account just a dimension.El microcrédito se presenta como una opción de financiamiento para estimular el sector productivo y apoyar el capital intelectual de la población que no tiene oportunidad de acceso a los créditos bancarios convencionales. A partir de la elaboración del mapa perceptual del consumidor de microcréditos de la ciudad de Bogotá, se llegó a la conclusión de que este valora tres dimensiones: capital de trabajo, permanecía y confianza, mientras que los individuos que no han adquirido crédito o que han empleado créditos bancarios solo valoran una dimensión fuertemente.

El mapa perceptual del consumidor de microcréditos para la ciudad de Bogotá

Microcredit typically refers to very small loans for unsalaried borrowers with little or no collateral and has been seen as a financing option to encourage the productive sector and support the brain capital of the population who has no typical bank loans. By mapping consumer microcredit perceptual in Bogota we could infer three dimensions projected: working capital, permanence and trust-worthiness, contrasting those people who do not have credit or any bank loans but take strongly into account just a dimension.El microcrédito se presenta como una opción de financiamiento para estimular el sector productivo y apoyar el capital intelectual de la población que no tiene oportunidad de acceso a los créditos bancarios convencionales. A partir de la elaboración del mapa perceptual del consumidor de microcréditos de la ciudad de Bogotá, se llegó a la conclusión de que este valora tres dimensiones: capital de trabajo, permanecía y confianza, mientras que los individuos que no han adquirido crédito o que han empleado créditos bancarios solo valoran una dimensión fuertemente.

El mapa perceptual del consumidor de microcréditos para la ciudad de Bogotá

El microcrédito se presenta como una opción de financiamiento para estimular el sector productivo y apoyar el capital intelectual de la población que no tiene oportunidad de acceso a los créditos bancarios convencionales. A partir de la elaboración del mapa perceptual del consumidor de microcréditos de la ciudad de Bogotá, se llegó a la conclusión de que este valora tres dimensiones: capital de trabajo, permanecía y confianza, mientras que los individuos que no han adquirido crédito o que han empleado créditos bancarios solo valoran una dimensión fuertemente.