Z*****N
发帖数: 112 | 1 Hi everyone,
I plan to make a point mutation (ACT->GCT) on the target gene and generate a
knock-in mice. I found out that ACT just locates exon-intron junction (e.g.[exon A]--intron--[CT exon]). Does the point mutation likely affect RNA
splicing? If so, any suggestion?
Many Thanks. |
Z*****N
发帖数: 112 | |
h****g
发帖数: 439 | 3 不知道，帮顶。PS，你真够Lucky的
a
g.[exon A]--intron--[CT exon]). Does the point mutation likely affect RNA
【在 Z*****N 的大作中提到】
: Hi everyone,
: I plan to make a point mutation (ACT->GCT) on the target gene and generate a
: knock-in mice. I found out that ACT just locates exon-intron junction (e.g.[exon A]--intron--[CT exon]). Does the point mutation likely affect RNA
: splicing? If so, any suggestion?
: Many Thanks.
|
H****N
发帖数: 997 | 4 not likely, but I would test it in cell culture first considering the effort
involved in generating a K/I mouse. |
J********3
发帖数: 3151 | 5 T变D或E啥的不行吗？实验证明非得T变A？
a
g.[exon A]--intron--[CT exon]). Does the point mutation likely affect RNA
【在 Z*****N 的大作中提到】
: Hi everyone,
: I plan to make a point mutation (ACT->GCT) on the target gene and generate a
: knock-in mice. I found out that ACT just locates exon-intron junction (e.g.[exon A]--intron--[CT exon]). Does the point mutation likely affect RNA
: splicing? If so, any suggestion?
: Many Thanks.
|
Z*****N
发帖数: 112 | 6 Unfortunately, T has to been muated to A |
n**b
发帖数: 23 | 7 only boundary sequence in Intron is nonexceptional (that is to say, intron
has to start with GT,and end with AG). As for the sequence in Exon, it
usually ends with AG, and starts with GT. So It will be OK to change A to G
for your case. But as HENBEN said, you'd better to test it first. |
Z*****N
发帖数: 112 | 8 Thank everyone. It sounds like the mutated genomic DNA should be test for
splicing before puttng into mice.
Is there any way to test the splicing in vitro or in vivo? |
i*********0
发帖数: 915 | 9 Normally I just leave the AG GT in the introns unchanged. I do not care
about the end and beginning of the exon.
whatever, when you have the targeted ES cells, you can detect it with RT-PCR.
G
【在 n**b 的大作中提到】
: only boundary sequence in Intron is nonexceptional (that is to say, intron
: has to start with GT,and end with AG). As for the sequence in Exon, it
: usually ends with AG, and starts with GT. So It will be OK to change A to G
: for your case. But as HENBEN said, you'd better to test it first.
|
