Is 1 kb of DNA strand good for CRISPR?

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Is there an azole specific efflux family in fungi?

It looks like azole transporters are similar in many fungi, saprophytes, plant and human pathogens. Many of hte azole transporters have homology to CDR1, like BcatrD (and a bit B) from Botrytis...

03 April 2013 1,314 1 View

What is a good website to compare lab products

As a technician I sometime want to compare different lab products. There are a lot of website and I am wondering which websites you use?

01 January 1970 5,205 13 View

Why am I getting 'Infinity' and 'NaN' in my mixed-mode energy release values?

Hi everyone, I am currently running a mixed-mode simulation of a DCB using a user-defined cohesive zone model (UEL), however, the results I am getting don't make sense. I used the same UEL for...

24 February 2021 4,157 1 View

In what ways do fellow reaerchers think that mRNA vaccination may be under CRISPR systems technology application ?

There are many ways for mRNA to intervene with genome expression. This question is raised for experts in CRISPR systems. Any RNA does not stand alone within acell. There are many ways to become...

18 February 2021 9,109 5 View

How can I completely delete a protein on the genomic level using Crispr?

I want to completely delete or largely truncate a gene and not only achieve a functional knockout in a cell line. I can only transfect my cells with the Crispr/Cas9 sgRNA RNP and not introduce a...

08 February 2021 2,032 2 View

Choise of cell line for neurobiology?

I am working with myelin-related proteins and interested to do cell experiments with my target protein in cells. I have created lentiviruses for my proteins, and mutants and also CRISPR set-up for...

02 February 2021 8,083 1 View

Does gene insertion via CRISPR/Cas9 result in chromosomal abnormalities?

Hello, I hope to insert a gene into mammalian cells in culture using CRISPR/Cas9 and an HDR template. Will insertion of a 1 kilobase gene cause chromosomal abnormalities and effect cell...

31 January 2021 2,516 1 View

Does anyone has a protocol for transfecting AML12 cells?

I am optimizing a CRISPR/CAS9 system in vitro before using it in vivo. I would like to transfect AML12 cells with my pAAV-sgRNA-vector to validate editing efficiency before generating virus.

26 January 2021 8,733 2 View

How to apply predefined temperature to generate prestressed force in Abaqus?

Dear researcher, I'm applying a thermal strain to generate the prestressed force in the steel strand of my concrete wall model using predefined temperature in Abaqus as shown in Figure 1. In...

26 January 2021 7,058 7 View

Which miRNA arm is a right target for analysis, 3p or 5p?

I'm working on selecting mouse miRNAs to analyze and stumbling upon one confusing issue, some of the papers do not specify which arm they analyzed and they just state something like " miR-34a was...

16 January 2021 3,792 4 View

Should I add a promoter upstream of my CRISPR-inserted gene?

Hello, I want to insert a gene into my cells using sgRNA, Cas9, and an HDR template. The gene that I will insert into has its own promoter. Because I will cut the gene downstream of its start...

14 January 2021 1,170 2 View

Does anyone have problems performing CRISPR KO in human primary T cells?

For the past few months, I have been troubleshooting a CRISPR KO of human CD3e in human primary T cells. I got the KO to work one time using 1250 ng TrueCut Cas9 (Thermo) + 250 ng (or 7.5 pmol)...

11 January 2021 6,016 3 View

Muhammad Ali

Please follow : https://academic.oup.com/nar/article/48/2/974/5651333Article Delivering CRISPR: a review of the challenges and approaches

Article Comparative analysis of single-stranded DNA donors to genera...

Roy Rabinowitz

Large insertions usually result in lower HDR rates. To my knowledge, the upper limit is around 2-2.5kb.